LOCUS       sequence-368742-        6622 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 6622)
  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 6622)
  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-PILRB-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6622
                     /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(2338..2357)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2370..2388)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2371..2388)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2598..2622)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2619..2822)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2648..2668)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2823..3202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3369..3385
                     /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(3841..3858)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3841..3857)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3956..3973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4127..4715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4207..4226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4801..4820)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4826..4845)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4887..4978
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4979..5047,5048..5839)
                     /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(5197..5216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5892..6013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5903..5922)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5957..5976
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6143..6622,1..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(6226..6246)
                     /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 cgcctctaat ggcggtatcc aggctcaggt gccagctctc
     1741 agagtgtccc ttgctctctg tcactctcag gccggcgatt gttgttgtgc tgctaggcct
     1801 ccaagttgtg gtggtggtca cggcctgggt gattgtcagc ttggtgccct tgatgctctg
     1861 cagctgctgt ctaccactcc gcctggtatc cagttccact ctgcagaagt agacgctctg
     1921 gtcctctttc cgcaggttgc tgattctcag gaagccgctc tcttggccct ctgtccagtt
     1981 caggaacagc cggttcacgt agtccttgtg gatgctaggt ggtcttgtgc tgtagaagct
     2041 ctggccgtgg aagtggcctc ttctccaaga gatccgcaca ttgggcacga tggcgagttc
     2101 ccaggggtag tagaagctga aggggatttc cacgctgccg cccatagagg cagacaggtg
     2161 cttaggctgt gtcacgccat acaggtagct agggccagag cctgtagatc caccaggttg
     2221 ggcggccgca gcgaacacca ggcagaagat gtagctcagg gcaatgatgg tcttcatggt
     2281 ggcgctagca ggcgcgtttt cgatagaacc gaggtgcagt tggacctggg agtggacacc
     2341 tgtggagaga aaggcaaagt ggatgttatt ctatagtgtc acctaaatcg tatgtgtatg
     2401 atacataagg ttatgtatta attgtagccg cgttctaacg aagccaaggg ggtgggccta
     2461 tagactctat aggcggtact tacgtcactc ttggcacggg gaatccgcgt tccaatgcac
     2521 cgttcccggc cgcggaggct ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt
     2581 ggatggcgtc tccaggcgat ctgacggttc actaaacgag ctctgcttat atagacctcc
     2641 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     2701 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     2761 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     2821 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     2881 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     2941 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     3001 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     3061 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     3121 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     3181 taactagtca ataatcaatg ccaacatggc ggtcatattg gacatgagcc aatataaatg
     3241 tacatattat gatatagata caacgtatgc aatggccaat agccaatatt gatttatgct
     3301 atataaccaa tgaataatat ggctaatggc caatattgag tcaacgccac tgtgcatgaa
     3361 gctcgtaatt gttatccgct cacaattaag ggcgacacaa aatttattct aaatgataat
     3421 aaatactgat aacatcttat agtttgtatt atattttgta ttatcgttga catgtataat
     3481 tttgatatca aaaactgatt ttccctttat tattttcgag atttattttc ttaattctct
     3541 ttaacaaact agaaatattg tatatacaaa aaatcataaa taatagatga atagtttaat
     3601 tataggtgtt catcaatcga aaaagcaacg tatcttattt aaagtgcgtt gcttttttct
     3661 catttataag gttaaataat tctcatatat caagcaaagt gacaggcgcc cttaaatatt
     3721 ctgacaaatg ctctttccct aaactccccc cataaaaaaa cccgccgaag cgggttttta
     3781 cgttatttgc ggattaacga ttactcgtta tcagaaccgc ccagggggcc cgagcttaag
     3841 actggccgtc gttttacaac acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3901 ggggccttct gcttagtttg atgcctggca gttccctact ctcgccttcc gcttcctcgc
     3961 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4021 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4081 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4141 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4201 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4261 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4321 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4381 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4441 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4501 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtgggctaac tacggctaca
     4561 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4621 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4681 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4741 ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt taagggattt tggtcatgag
     4801 caaggggtgt tatgagccat attcaggtat aaatgggctc gcgataatgt tcagaattgg
     4861 ttaattggtt gtaacactga cccctatttg tttatttttc taaatacatt caaatatgta
     4921 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatat
     4981 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5041 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5101 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5161 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5221 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5281 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5341 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5401 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5461 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5521 tgtagcgatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5581 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5641 ggcccttccg gctggctggt ttattgctga taaatccgga gccggtgagc gtggttctcg
     5701 cggtatcatc gcagcgctgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5761 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5821 actgattaag cattggtaat gaccatgatt acgccaagct ctagctagag gtcgacggta
     5881 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     5941 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     6001 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     6061 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     6121 aatcgaaatc tcgtagcacg tgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg
     6181 gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga
     6241 ggttgcgggg cgcggatgtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca
     6301 ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga
     6361 cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct
     6421 gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga
     6481 acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt
     6541 ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct
     6601 cggtgacccg ctcgatgtgg cg
//
