LOCUS       sequence-432345-        7114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            Fc(DAPA)-Avi-6xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7114)
  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 7114)
  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-KIR3DX1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7114
                     /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             2690..2734
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2735..2752
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2892..2909)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2898..3009
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3127..3147)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3132..3461
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3312..3447
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3374..3393
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3509..3556
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3575..4174
                     /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(3575..3594)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4071..4091
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4304..4425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4341..4360)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4395..4414
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4478..5269,5270..5338))
                     /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     5101..5120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5339..5430)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5472..5491
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5497..5516
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5602..6190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6091..6110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6344..6361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6459..6476
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6460..6476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6932..6948
                     /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 cccagaaaat ttgtcctcat gccggcgctc aggacaagtt
     1021 ctctctgtct gcttggccct ctcctgtggt gcctcttgga ggaagagtga ccctgagctg
     1081 tcacagccac ctgagattcg tgatctggac catcttccag accaccggca caagaagcca
     1141 cgagctgcac acaggcctga gcaacaacat cacaatcagc cccgtgacac ctgagcacgc
     1201 cggcacatat agatgcgtgg gcatctacaa gcacgccagc aagtggtccg ccgagagcaa
     1261 ctctctgaag atcatcgtga ccggcctgtt caccaagcct agcatctctg cccatcctag
     1321 cagcctggtt cacgctggtg ctagggtgtc cctgagatgt catagcgagc tggccttcga
     1381 cgagttcatc ctgtacaaag agggccacat ccagcacagc cagcagctgg atcaaggcat
     1441 ggaagccggc atccactacg tggaagccgt gttttctatg ggccctgtga caccagctca
     1501 cgctggcgcc tatagatgct gcggctgttt cagccacagc agatacgagt ggtctgcccc
     1561 tagcgaccct ctggatatcg tgatcaccgg caagtataag aagcccagcc tgagcaccca
     1621 ggtggaccct atgatgagac tgggcgagaa gctgaccctg ttctgcagca gcgagatcag
     1681 cttcgaccag taccacctgt ttagacacgg cgtggcacac ggacaatggc tgtctggtgg
     1741 acagagacac agagaggcct tccaggccaa tttcagcgtg ggcagagcta cacctgtgcc
     1801 tggcggcacc tacagatgct acggcagctt caacgacagc ccctacaagc ctcctgtgac
     1861 cagatgcaac ttcacccctc aagagacact gcgggtgctg ctgtgtcact cccagaatcc
     1921 tccactgaat ctgacccatc tggccctgaa ggatagcccc gccacatgta tctgcagcct
     1981 ggactctcaa ggcgcgccgg gcggcgacaa gacccacacc tgccccccct gcccagctcc
     2041 agaactgctg ggagggcctt ccgtgttcct gttccccccc aagcccaagg acaccctgat
     2101 gatcagcagg acccccgagg tgacctgcgt ggtggtggcc gtgtcccacg aggacccaga
     2161 ggtgaagttc aactggtacg tggacggcgt ggaggtgcac aacgccaaga ccaagcccag
     2221 agaggagcag tacaacagca cctacagggt ggtgtccgtg ctgaccgtgc tgcaccagga
     2281 ctggctgaac ggcaaagaat acaagtgcaa agtctccaac aaggccctgg ctgccccaat
     2341 cgaaaagaca atcagcaagg ccaagggcca gccacgggag ccccaggtgt acaccctgcc
     2401 ccccagccgg gaggagatga ccaagaacca ggtgtccctg acctgtctgg tgaagggctt
     2461 ctaccccagc gatatcgccg tggagtggga gagcaacggc cagcccgaga acaactacaa
     2521 gaccaccccc ccagtgctgg acagcgacgg cagcttcttc ctgtacagca agctgaccgt
     2581 ggacaagtcc aggtggcagc agggcaacgt gttcagctgc agcgtgatgc acgaggccct
     2641 gcacaaccac tacacccaga agtccctgag cctgagcccc ggcggcagcg gtcttaatga
     2701 tattttcgag gctcagaaga ttgagtggca tgaacaccac caccatcacc actaaggccg
     2761 gcctgtagtc actggcacaa cgcgttaccc ggtaagccaa tcgggtatac acggtcgtca
     2821 tactgcagac agggttcttc tactttgcaa gatagtcttg agtagtaaaa taaatagata
     2881 gagaaaatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2941 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3001 aattgcatca caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3061 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3121 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3181 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3301 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3361 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3421 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3481 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     3541 catagtataa tacgacaagg tgaggaacta aaccatgacc gagtacaagc ccacggtgcg
     3601 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3661 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3721 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3781 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     3841 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     3901 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     3961 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4021 ggcggccgag cgcgccgggg tgcccgcctt cctggagaca tccgcgcccc gcaacctccc
     4081 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4141 cacctggtgc atgacccgca agcccggtgc ctgacacgtg ctacgagatt tcgattccac
     4201 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4261 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4321 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4381 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4441 gtcgacctct agctagagct tggcgtaatc atggtcatta ccaatgctta atcagtgagg
     4501 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4561 agataactac gatacgggag ggcttaccat ctggccccag cgctgcgatg ataccgcgag
     4621 aaccacgctc accggctccg gatttatcag caataaacca gccagccgga agggccgagc
     4681 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4741 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatc gctacaggca
     4801 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4861 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4921 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4981 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5041 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5101 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5161 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5221 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5281 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatat
     5341 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5401 tatttgaatg tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa
     5461 ttctgaacat tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgctca
     5521 tgaccaaaat cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt
     5581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5761 gccgtagtta gcccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6361 ggaagcggaa ggcgagagta gggaactgcc aggcatcaaa ctaagcagaa ggcccctgac
     6421 ggatggcctt tttgcgtttc tacaaactct ttctgtgttg taaaacgacg gccagtctta
     6481 agctcgggcc ccctgggcgg ttctgataac gagtaatcgt taatccgcaa ataacgtaaa
     6541 aacccgcttc ggcgggtttt tttatggggg gagtttaggg aaagagcatt tgtcagaata
     6601 tttaagggcg cctgtcactt tgcttgatat atgagaatta tttaacctta taaatgagaa
     6661 aaaagcaacg cactttaaat aagatacgtt gctttttcga ttgatgaaca cctataatta
     6721 aactattcat ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag
     6781 aattaagaaa ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta
     6841 tacatgtcaa cgataataca aaatataata caaactataa gatgttatca gtatttatta
     6901 tcatttagaa taaattttgt gtcgccctta attgtgagcg gataacaatt acgagcttca
     6961 tgcacagtgg cgttgactca atattggcca ttagccatat tattcattgg ttatatagca
     7021 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     7081 tatattggct catgtccaat atgaccgcca tgtt
//
