LOCUS       sequence-410017-        6778 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 6778)
  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 6778)
  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-KIR2DL2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6778
                     /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             2354..2398
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2399..2416
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2556..2573)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2562..2673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2791..2811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2796..3125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2976..3111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3038..3057
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3173..3220
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3239..3838
                     /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(3239..3258)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3735..3755
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3968..4089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4005..4024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4059..4078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4142..4933,4934..5002))
                     /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     4765..4784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5003..5094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5136..5155
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5161..5180
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5266..5854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5755..5774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6008..6025
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6123..6140
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6124..6140
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6596..6612
                     /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 cccatgaagg cgtgcacaga aaaccttctc tgctggccca
     1021 tcctggccgg ctggttaagt ctgaagagac agtgatcctg cagtgttgga gcgacgtccg
     1081 cttcgagcac tttctgctgc acagagaggg caagttcaag gacacactgc acctgatcgg
     1141 agagcaccac gatggcgtgt ccaaggccaa tttcagcatc ggccccatga tgcaggatct
     1201 ggccggcaca tacagatgct acggctctgt gacacacagc ccctatcagc tgtctgcccc
     1261 tagcgatccc ctggatatcg tgatcaccgg cctgtacgag aagcctagcc tgtctgctca
     1321 gcctggacct acagttctgg ctggcgaaag cgtgaccctg agctgtagca gcagatccag
     1381 ctacgacatg taccacctgt ccagagaagg cgaggcccac gagtgtagat tcagcgccgg
     1441 acctaaagtg aacggcacct tccaggccga ttttccactg ggacctgcca cacacggcgg
     1501 cacctataga tgctttggca gcttcagaga cagcccttac gagtggtcca acagcagcga
     1561 tcctctgctg gtgtccgtga tcggcaaccc tagcaatagc tggccttctc caaccgagcc
     1621 tagcagcaag accggcaatc ccagacatct gcatggcgcg ccgggcggcg acaagaccca
     1681 cacctgcccc ccctgcccag ctccagaact gctgggaggg ccttccgtgt tcctgttccc
     1741 ccccaagccc aaggacaccc tgatgatcag caggaccccc gaggtgacct gcgtggtggt
     1801 ggccgtgtcc cacgaggacc cagaggtgaa gttcaactgg tacgtggacg gcgtggaggt
     1861 gcacaacgcc aagaccaagc ccagagagga gcagtacaac agcacctaca gggtggtgtc
     1921 cgtgctgacc gtgctgcacc aggactggct gaacggcaaa gaatacaagt gcaaagtctc
     1981 caacaaggcc ctggctgccc caatcgaaaa gacaatcagc aaggccaagg gccagccacg
     2041 ggagccccag gtgtacaccc tgccccccag ccgggaggag atgaccaaga accaggtgtc
     2101 cctgacctgt ctggtgaagg gcttctaccc cagcgatatc gccgtggagt gggagagcaa
     2161 cggccagccc gagaacaact acaagaccac ccccccagtg ctggacagcg acggcagctt
     2221 cttcctgtac agcaagctga ccgtggacaa gtccaggtgg cagcagggca acgtgttcag
     2281 ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc cagaagtccc tgagcctgag
     2341 ccccggcggc agcggtctta atgatatttt cgaggctcag aagattgagt ggcatgaaca
     2401 ccaccaccat caccactaag gccggcctgt agtcactggc acaacgcgtt acccggtaag
     2461 ccaatcgggt atacacggtc gtcatactgc agacagggtt cttctacttt gcaagatagt
     2521 cttgagtagt aaaataaata gatagagaaa atcagcctcg actgtgcctt ctagttgcca
     2581 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2641 tgtcctttcc taataaaatg aggaaattgc atcacaacac tcaaccctat ctcggtctat
     2701 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2761 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2941 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3001 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3061 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3121 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     3181 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     3241 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     3301 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     3361 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     3421 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     3481 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     3541 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     3601 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     3661 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     3721 gacatccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     3781 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaca
     3841 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3901 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3961 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4021 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4081 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4141 attaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4201 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4261 ccagcgctgc gatgataccg cgagaaccac gctcaccggc tccggattta tcagcaataa
     4321 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4381 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4441 acgttgttgc catcgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4501 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4561 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4621 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4681 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4741 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4801 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4861 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4921 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4981 cacggaaatg ttgaatactc atattcttcc tttttcaata ttattgaagc atttatcagg
     5041 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5101 tcagtgttac aaccaattaa ccaattctga acattatcgc gagcccattt atacctgaat
     5161 atggctcata acaccccttg ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
     5221 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5281 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5341 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5401 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
     5461 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5521 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5581 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5641 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5701 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5761 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5821 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5881 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5941 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6001 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
     6061 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
     6121 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     6181 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     6241 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     6301 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     6361 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     6421 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     6481 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     6541 ataagatgtt atcagtattt attatcattt agaataaatt ttgtgtcgcc cttaattgtg
     6601 agcggataac aattacgagc ttcatgcaca gtggcgttga ctcaatattg gccattagcc
     6661 atattattca ttggttatat agcataaatc aatattggct attggccatt gcatacgttg
     6721 tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc gccatgtt
//
