LOCUS       sequence-396956-        7063 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 7063)
  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 7063)
  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-KIR3DL1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7063
                     /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             2639..2683
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2684..2701
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2841..2858)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2847..2958
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3076..3096)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3081..3410
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3261..3396
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3323..3342
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3458..3505
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3524..4123
                     /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(3524..3543)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4020..4040
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4253..4374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4290..4309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4344..4363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4427..5218,5219..5287))
                     /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     5050..5069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5288..5379)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5421..5440
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5446..5465
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5551..6139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6040..6059
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6293..6310
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6408..6425
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6409..6425
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6881..6897
                     /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 cccacatggg aggacaagat aagccttttc tgagcgcctg
     1021 gcctagcgct gttgttccta gaggtggaca cgtgaccctg cggtgccact acagacaccg
     1081 gttcaacaac ttcatgctgt acaaagagga ccggattcac atccccatct tccacggccg
     1141 gatcttccaa gagtccttca acatgagccc cgtgacaaca gcccacgccg gcaattacac
     1201 ctgtagaggc tctcaccctc acagccctac aggatggtcc gctcctagca accctgtggt
     1261 catcatggtc accggcaacc acagaaagcc tagcctgctt gcccatcctg gacctctggt
     1321 taagtctggc gagcgcgtta tcctgcagtg ttggagcgat attatgttcg agcacttctt
     1381 tctgcacaaa gagggcatca gcaaggaccc cagcagactc gtgggacaga ttcacgatgg
     1441 cgtgtccaag gccaacttca gcatcggacc tatgatgctg gccctggccg gcacatacag
     1501 atgttacggc tctgtgaccc acacacctta ccagctgtct gcccctagcg accctctgga
     1561 tatcgtcgtg acaggccctt acgagaagcc aagcctgtct gctcagccag gacctaaagt
     1621 gcaggccggc gaaagcgtga cactgagctg tagcagcaga tccagctacg acatgtacca
     1681 cctgagcaga gaaggcggag cccatgagag aaggctgcct gctgtgcgga aagtgaacag
     1741 aaccttccag gccgactttc ctctgggacc tgctacacac ggcggcacct atagatgctt
     1801 cggcagcttc agacacagcc cctacgagtg gtccgatcca tctgatcctc tgctggtgtc
     1861 cgtgaccggc aatccttcta gcagctggcc ttctccaacc gagcctagca gcaagagcgg
     1921 caaccccaga catctgcatg gcgcgccggg cggcgacaag acccacacct gccccccctg
     1981 cccagctcca gaactgctgg gagggccttc cgtgttcctg ttccccccca agcccaagga
     2041 caccctgatg atcagcagga cccccgaggt gacctgcgtg gtggtggccg tgtcccacga
     2101 ggacccagag gtgaagttca actggtacgt ggacggcgtg gaggtgcaca acgccaagac
     2161 caagcccaga gaggagcagt acaacagcac ctacagggtg gtgtccgtgc tgaccgtgct
     2221 gcaccaggac tggctgaacg gcaaagaata caagtgcaaa gtctccaaca aggccctggc
     2281 tgccccaatc gaaaagacaa tcagcaaggc caagggccag ccacgggagc cccaggtgta
     2341 caccctgccc cccagccggg aggagatgac caagaaccag gtgtccctga cctgtctggt
     2401 gaagggcttc taccccagcg atatcgccgt ggagtgggag agcaacggcc agcccgagaa
     2461 caactacaag accacccccc cagtgctgga cagcgacggc agcttcttcc tgtacagcaa
     2521 gctgaccgtg gacaagtcca ggtggcagca gggcaacgtg ttcagctgca gcgtgatgca
     2581 cgaggccctg cacaaccact acacccagaa gtccctgagc ctgagccccg gcggcagcgg
     2641 tcttaatgat attttcgagg ctcagaagat tgagtggcat gaacaccacc accatcacca
     2701 ctaaggccgg cctgtagtca ctggcacaac gcgttacccg gtaagccaat cgggtataca
     2761 cggtcgtcat actgcagaca gggttcttct actttgcaag atagtcttga gtagtaaaat
     2821 aaatagatag agaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2881 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2941 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     3001 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3061 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3241 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3301 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3361 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3421 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     3481 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc
     3541 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     3601 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     3661 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     3721 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     3781 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     3841 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     3901 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     3961 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg
     4021 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4081 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt
     4141 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4201 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4261 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4321 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4381 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa
     4441 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4501 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga
     4561 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa
     4621 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4681 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     4741 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4801 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4861 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4921 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4981 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5041 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5101 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5161 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5221 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5281 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5341 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca
     5401 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc
     5461 ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     5521 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5581 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5641 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5701 tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct
     5761 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5821 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5881 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5941 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6001 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6061 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6121 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6181 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6241 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6301 agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag
     6361 gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg
     6421 ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa
     6481 taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt
     6541 gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat
     6601 aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac
     6661 ctataattaa actattcatc tattatttat gattttttgt atatacaata tttctagttt
     6721 gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata
     6781 tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag atgttatcag
     6841 tatttattat catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta
     6901 cgagcttcat gcacagtggc gttgactcaa tattggccat tagccatatt attcattggt
     6961 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     7021 atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
