LOCUS       sequence-387050-        7572 bp    DNA     linear   UNA 14-APR-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      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7572)
  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 7572)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-SIGLEC10-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7572
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    59..75
                     /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)."
     enhancer        242..621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        622..825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     776..796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     822..846
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1056..1074
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1056..1073
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1087..1106
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1218..1227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3525..3569
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3570..3587
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3727..3744)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3733..3844
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3962..3982)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3967..4296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4147..4282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4209..4228
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4344..4391
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4410..5009
                     /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(4410..4429)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4906..4926
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5139..5260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5176..5195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5230..5249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5313..6104,6105..6173))
                     /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     5936..5955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6174..6265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6307..6326
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6332..6351
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6437..7025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6926..6945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7179..7196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7294..7311
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7295..7311
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 rctataagat gttatcagta tttattatca tttagaataa attttgtgtc gcccttaatt
       61 gtgagcggat aacaattacg agcttcatgc acagtggcgt tgactcaata ttggccatta
      121 gccatattat tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg
      181 ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg accgccatgt
      241 tggcattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
      841 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
      901 tccagcctcc gcggccggga acggtgcatt ggaacgcgga ttccccgtgc caagagtgac
      961 gtaagtaccg cctatagagt ctataggccc acccccttgg cttcgttaga acgcggctac
     1021 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaataaca
     1081 tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcac ctcggttcta
     1141 tcgaaaacgc gcctgctagc gccaccatga agaccatcat tgccctgagc tacatcttct
     1201 gcctggtgtt cgctgcggcc gccatggatg gcagattctg gatcagagtg caagagtccg
     1261 tgatggtgcc tgagggcctg tgtatcagcg tgccctgcag cttcagctac cccagacagg
     1321 attggacagg cagcacacca gcctacggct actggtttaa ggccgtgacc gaaaccacca
     1381 agggcgctcc agtggccaca aaccaccaga gcagagaagt ggaaatgagc acccggggca
     1441 gattccagct gacaggcgat cctgccaagg gcaattgcag cctggtcatc agagatgccc
     1501 agatgcagga cgagagccag tacttcttca gagtggaacg gggcagctac gtgcggtaca
     1561 actttatgaa cgacggattc ttcctgaaag tgacggccct gacacagaaa cccgacgtgt
     1621 acatccccga gacactggaa cctggccagc ctgtgacagt gatctgcgtg ttcaactggg
     1681 ccttcgagga atgccctcca ccttcctttt cttggactgg cgccgctctg tctagccagg
     1741 gcacaaagcc taccaccagc cactttagcg tgctgagctt cacccctaga cctcaggacc
     1801 acaacaccga cctgacctgc cacgtggact tcagcagaaa aggcgtgtca gcccagagaa
     1861 ccgtgcggct gagagtggcc tatgctccca gagatctcgt gatcagcatc agccgggaca
     1921 acacccctgc tctggaacct cagcctcagg gcaatgtgcc ttacctggaa gcccagaagg
     1981 gccagttcct gagactgctg tgtgccgccg attctcagcc tcctgctaca ctgtcttggg
     2041 tgctgcagaa cagagtgctg agcagctctc acccttgggg ccctagacca ctgggacttg
     2101 aactgcctgg tgtcaaggcc ggcgacagcg gcagatacac atgcagagcc gagaacagac
     2161 tgggcagcca gcagagagca ctggatctga gcgtgcagta ccctcctgag aacctgagag
     2221 tgatggtgtc ccaggccaac cggaccgtgc tggaaaatct cggcaatggc acaagcctgc
     2281 ctgtgctgga aggccagtct ctgtgtctcg tgtgtgtgac ccacagcagc cctccagcca
     2341 gactctcttg gacacagcga ggacaggtgc tgtctcctag ccagccttct gatcctggcg
     2401 tcctggaact gcctagagtg caggttgagc acgagggcga gttcacctgt cacgccagac
     2461 atcctctggg ctctcagcac gtttccctga gcctgtctgt gcactacagc cctaagctgc
     2521 tgggccctag ctgttcttgg gaagccgaag gcctgcactg cagctgtagc tctcaagcct
     2581 ctcctgctcc tagcctgaga tggtggctgg gcgaagaact gctcgagggc aatagctccc
     2641 aggacagctt cgaagtgacc ccttcttctg ccggaccttg ggccaatagc tctctgtctc
     2701 tgcacggcgg actgagcagt ggactgagac tgagatgcga ggcctggaat gtgcacggtg
     2761 ctcagtctgg ctctatcctg cagctgcccg ataagaaggg cctgatcagc accgccttca
     2821 gcaatggcgc gccgggcggc gacaagaccc acacctgccc cccctgccca gctccagaac
     2881 tgctgggagg gccttccgtg ttcctgttcc cccccaagcc caaggacacc ctgatgatca
     2941 gcaggacccc cgaggtgacc tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga
     3001 agttcaactg gtacgtggac ggcgtggagg tgcacaacgc caagaccaag cccagagagg
     3061 agcagtacaa cagcacctac agggtggtgt ccgtgctgac cgtgctgcac caggactggc
     3121 tgaacggcaa agaatacaag tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa
     3181 agacaatcag caaggccaag ggccagccac gggagcccca ggtgtacacc ctgcccccca
     3241 gccgggagga gatgaccaag aaccaggtgt ccctgacctg tctggtgaag ggcttctacc
     3301 ccagcgatat cgccgtggag tgggagagca acggccagcc cgagaacaac tacaagacca
     3361 cccccccagt gctggacagc gacggcagct tcttcctgta cagcaagctg accgtggaca
     3421 agtccaggtg gcagcagggc aacgtgttca gctgcagcgt gatgcacgag gccctgcaca
     3481 accactacac ccagaagtcc ctgagcctga gccccggcgg cagcggtctt aatgatattt
     3541 tcgaggctca gaagattgag tggcatgaac accaccacca tcaccactaa ggccggcctg
     3601 tagtcactgg cacaacgcgt tacccggtaa gccaatcggg tatacacggt cgtcatactg
     3661 cagacagggt tcttctactt tgcaagatag tcttgagtag taaaataaat agatagagaa
     3721 aatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3781 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3841 catcacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3901 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3961 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4021 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4081 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4141 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4201 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4261 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4321 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     4381 tataatacga caaggtgagg aactaaacca tgaccgagta caagcccacg gtgcgcctcg
     4441 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4501 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4561 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4621 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4681 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4741 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4801 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4861 ccgagcgcgc cggggtgccc gccttcctgg agacatccgc gccccgcaac ctccccttct
     4921 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4981 ggtgcatgac ccgcaagccc ggtgcctgac acgtgctacg agatttcgat tccaccgccg
     5041 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5101 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5161 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5221 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5281 cctctagcta gagcttggcg taatcatggt cattaccaat gcttaatcag tgaggcacct
     5341 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5401 actacgatac gggagggctt accatctggc cccagcgctg cgatgatacc gcgagaacca
     5461 cgctcaccgg ctccggattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5521 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5581 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg
     5641 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5701 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5761 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5821 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5881 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5941 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6001 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6061 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6121 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catattcttc
     6181 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6241 gaatgtattt agaaaaataa acaaataggg gtcagtgtta caaccaatta accaattctg
     6301 aacattatcg cgagcccatt tatacctgaa tatggctcat aacacccctt gctcatgacc
     6361 aaaatccctt aacgtgagtt acgcgcgcgt cgttccactg agcgtcagac cccgtagaaa
     6421 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6481 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6541 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6601 agttagccca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6661 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6721 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6781 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6841 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6901 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6961 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7021 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7081 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7141 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7201 cggaaggcga gagtagggaa ctgccaggca tcaaactaag cagaaggccc ctgacggatg
     7261 gcctttttgc gtttctacaa actctttctg tgttgtaaaa cgacggccag tcttaagctc
     7321 gggccccctg ggcggttctg ataacgagta atcgttaatc cgcaaataac gtaaaaaccc
     7381 gcttcggcgg gtttttttat ggggggagtt tagggaaaga gcatttgtca gaatatttaa
     7441 gggcgcctgt cactttgctt gatatatgag aattatttaa ccttataaat gagaaaaaag
     7501 caacgcactt taaataagat acgttgcttt ttcgattgat gaacacctat aattmaacta
     7561 ttcatctatt ay
//
