LOCUS       sequence-401534-        7774 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 7774)
  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 7774)
  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-FCRL3-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7774
                     /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             3350..3394
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3395..3412
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3552..3569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3558..3669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3787..3807)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3792..4121
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3972..4107
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4034..4053
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4169..4216
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4235..4834
                     /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(4235..4254)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4731..4751
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4964..5085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5001..5020)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5055..5074
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5138..5929,5930..5998))
                     /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     5761..5780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5999..6090)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6132..6151
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6157..6176
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6262..6850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6751..6770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7004..7021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7119..7136
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7120..7136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7592..7608
                     /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 ccggtgttgc tcctaaagct gttctgctgc tgaaccctcc
     1021 ttggagcaca gcctttaagg gcgagaaggt ggccctgatc tgtagctcta tcagccattc
     1081 tctggcccag ggcgacacct actggtatca cgatgagaag ctgctgaaga tcaagcacga
     1141 caagatccag atcaccgagc ctggcaacta ccagtgcaag acaagaggca gcagcctgtc
     1201 tgatgccgtg cacgtggaat tcagccccga ttggctgatt ctgcaggctc tgcaccctgt
     1261 gttcgagggc gacaatgtga tcctgagatg ccagggcaaa gacaacaaga acacccacca
     1321 gaaggtgtac tacaaggacg gcaagcagct gcccaacagc tacaacctgg aaaagatcac
     1381 cgtgaacagc gtgtccagag acaacagcaa gtaccactgc accgcctacc ggaagttcta
     1441 catcctggac atcgaagtga ccagcaagcc cctgaacatc caggtgcaag agctgtttct
     1501 gcaccccgtg ctgagagcca gcagctctac acctattgag ggcagcccta tgacactgac
     1561 ctgcgagaca cagctgagcc ctcaaagacc tgacgtccag ctgcagttca gcctgttcag
     1621 agatagccag acactcggcc tcggctggtc cagatctcct agactgcaga tccctgccat
     1681 gtggacagag gacagcggct cttattggtg cgaggtggaa accgtgacac acagcatcaa
     1741 gaagcggagc ctgcggagcc agattagagt gcagagggtg cccgtgtcca acgtgaacct
     1801 cgaaatcaga cctacaggcg gccagctgat cgaaggcgag aatatggtgc tgatctgcag
     1861 cgtggcccaa ggctctggca ccgtgacatt ctcttggcac aaagagggca gagtcagaag
     1921 cctgggcaga aagacacaga gaagcctgct ggccgaactg cacgtgctga cagtgaaaga
     1981 aagcgacgcc ggaaggtact actgtgccgc cgataatgtg cacagcccca tcctgagcac
     2041 atggatcaga gtgaccgtgc gcatccctgt gtctcatcct gtgctgacct tcagagcccc
     2101 tagagcacac acagttgtgg gcgatctgct ggaactgcac tgcgagtctc tgagaggcag
     2161 cccacctatc ctgtaccggt tctaccacga ggacgtgacc ctgggcaatt cttctgctcc
     2221 ttctggcgga ggcgccagct tcaatctgtc tctgacagcc gagcacagcg gcaattacag
     2281 ctgtgatgct gataacggcc tgggagccca gcattctcat ggcgttagtc tgagagtgac
     2341 agtgcctgtg tccagaccag tgctgacatt gagagcacct ggcgctcagg ccgttgttgg
     2401 agatctgctc gagcttcatt gcgagagcct gaggggcagc ttccccattc tgtactggtt
     2461 ttatcatgag gacgacaccc tgggaaacat cagcgctcat agcggaggcg gagcctcttt
     2521 caatctgagc ctgacaacag agcactccgg caactactct tgcgaggctg ataatggact
     2581 cggcgctcag cacagcaagg tggtcacact gaatgtgacc ggcacctcca gaaaccggac
     2641 aggccttaca ggcgcgccgg gcggcgacaa gacccacacc tgccccccct gcccagctcc
     2701 agaactgctg ggagggcctt ccgtgttcct gttccccccc aagcccaagg acaccctgat
     2761 gatcagcagg acccccgagg tgacctgcgt ggtggtggcc gtgtcccacg aggacccaga
     2821 ggtgaagttc aactggtacg tggacggcgt ggaggtgcac aacgccaaga ccaagcccag
     2881 agaggagcag tacaacagca cctacagggt ggtgtccgtg ctgaccgtgc tgcaccagga
     2941 ctggctgaac ggcaaagaat acaagtgcaa agtctccaac aaggccctgg ctgccccaat
     3001 cgaaaagaca atcagcaagg ccaagggcca gccacgggag ccccaggtgt acaccctgcc
     3061 ccccagccgg gaggagatga ccaagaacca ggtgtccctg acctgtctgg tgaagggctt
     3121 ctaccccagc gatatcgccg tggagtggga gagcaacggc cagcccgaga acaactacaa
     3181 gaccaccccc ccagtgctgg acagcgacgg cagcttcttc ctgtacagca agctgaccgt
     3241 ggacaagtcc aggtggcagc agggcaacgt gttcagctgc agcgtgatgc acgaggccct
     3301 gcacaaccac tacacccaga agtccctgag cctgagcccc ggcggcagcg gtcttaatga
     3361 tattttcgag gctcagaaga ttgagtggca tgaacaccac caccatcacc actaaggccg
     3421 gcctgtagtc actggcacaa cgcgttaccc ggtaagccaa tcgggtatac acggtcgtca
     3481 tactgcagac agggttcttc tactttgcaa gatagtcttg agtagtaaaa taaatagata
     3541 gagaaaatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3601 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3661 aattgcatca caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3721 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3781 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3841 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3901 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3961 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4021 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4081 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4141 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     4201 catagtataa tacgacaagg tgaggaacta aaccatgacc gagtacaagc ccacggtgcg
     4261 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4321 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4381 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4441 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4501 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4561 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4621 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4681 ggcggccgag cgcgccgggg tgcccgcctt cctggagaca tccgcgcccc gcaacctccc
     4741 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4801 cacctggtgc atgacccgca agcccggtgc ctgacacgtg ctacgagatt tcgattccac
     4861 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4921 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4981 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5041 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5101 gtcgacctct agctagagct tggcgtaatc atggtcatta ccaatgctta atcagtgagg
     5161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5221 agataactac gatacgggag ggcttaccat ctggccccag cgctgcgatg ataccgcgag
     5281 aaccacgctc accggctccg gatttatcag caataaacca gccagccgga agggccgagc
     5341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatc gctacaggca
     5461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatat
     6001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6061 tatttgaatg tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa
     6121 ttctgaacat tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgctca
     6181 tgaccaaaat cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt
     6241 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6301 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6361 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6421 gccgtagtta gcccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6481 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6541 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6601 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6661 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6721 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6781 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6841 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6901 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6961 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7021 ggaagcggaa ggcgagagta gggaactgcc aggcatcaaa ctaagcagaa ggcccctgac
     7081 ggatggcctt tttgcgtttc tacaaactct ttctgtgttg taaaacgacg gccagtctta
     7141 agctcgggcc ccctgggcgg ttctgataac gagtaatcgt taatccgcaa ataacgtaaa
     7201 aacccgcttc ggcgggtttt tttatggggg gagtttaggg aaagagcatt tgtcagaata
     7261 tttaagggcg cctgtcactt tgcttgatat atgagaatta tttaacctta taaatgagaa
     7321 aaaagcaacg cactttaaat aagatacgtt gctttttcga ttgatgaaca cctataatta
     7381 aactattcat ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag
     7441 aattaagaaa ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta
     7501 tacatgtcaa cgataataca aaatataata caaactataa gatgttatca gtatttatta
     7561 tcatttagaa taaattttgt gtcgccctta attgtgagcg gataacaatt acgagcttca
     7621 tgcacagtgg cgttgactca atattggcca ttagccatat tattcattgg ttatatagca
     7681 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     7741 tatattggct catgtccaat atgaccgcca tgtt
//
