LOCUS       sequence-409884-        7402 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 7402)
  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 7402)
  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-BTNL2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7402
                     /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             2978..3022
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3023..3040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3180..3197)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3186..3297
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3415..3435)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3420..3749
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3600..3735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3662..3681
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3797..3844
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3863..4462
                     /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(3863..3882)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4359..4379
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4592..4713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4629..4648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4683..4702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4766..5557,5558..5626))
                     /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     5389..5408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5627..5718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5760..5779
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5785..5804
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5890..6478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6379..6398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6632..6649
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6747..6764
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6748..6764
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7220..7236
                     /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 ccaaacagag cgaggacttc agagtgatcg gccctgctca
     1021 ccctattctg gctggcgtgg gagaagatgc cctgctgact tgtcagctgc tgcccaagag
     1081 aaccaccatg cacgtggaag tgcggtggta cagaagcgag cctagcacac ctgtgttcgt
     1141 gcacagagat ggcgtggaag tcaccgagat gcagatggaa gagtacagag gctgggtcga
     1201 gtggatcgag aacggaatcg ccaagggcaa cgtggccctg aagatccaca acatccagcc
     1261 tagcgacaac ggccagtatt ggtgccactt ccaagacggc aactactgcg gcgagacaag
     1321 cctgctgctg aaagtggctg gactgggctc tgcccctagc atccacatgg aaggacctgg
     1381 cgaatctggc gtgcagctcg tgtgtacagc cagaggatgg ttccctgagc ctcaggtgta
     1441 ctgggaagat atccggggcg agaagctgct ggccgtgtct gagcacagaa tccaggacaa
     1501 ggacggcctg ttctacgccg aggctacact ggtcgtgcgg aatgcctctg ccgagtccgt
     1561 gtcttgcctg gtgcataacc ctgtgctgac cgaagagaag ggcagcgtga tcagcctgcc
     1621 tgagaaactg cagaccgagc tggcctctct gaaagtgaat ggccccagcc agcctatcct
     1681 cgtcagagtg ggcgaagata ttcagctgac ctgctatctg agccccaagg ccaacgctca
     1741 gagcatggaa gtccgctggg accgcagcca tagatatcct gccgtgcacg tgtacatgga
     1801 cggcgatcat gttgccggcg agcagatggc cgaataccgg ggaagaacag tgctggtgtc
     1861 cgacgccatt gacgagggca gactgacact gcagatcctg agcgccagac catccgacga
     1921 tggccagtac agatgcctgt tcgagaagga cgacgtgtac caagaagcca gcctggacct
     1981 gaaggtggtg tccctgggaa gcagccctct gatcacagtg gaaggccaag aggacggcga
     2041 aatgcagcct atgtgtagca gcgacggctg gttcccacag cctcatgtgc cttggagaga
     2101 tatggaaggc aagactatcc ccagcagcag ccaggctctg acacagggat ctcacggact
     2161 gtttcacgtg cagaccctgc tgagagtgac caacatcagc gccgtggatg tgacctgctc
     2221 tatcagcatc ccattcctcg gagaggaaaa gatcgccacc tttagcctga gcggctgggg
     2281 cgcgccgggc ggcgacaaga cccacacctg ccccccctgc ccagctccag aactgctggg
     2341 agggccttcc gtgttcctgt tcccccccaa gcccaaggac accctgatga tcagcaggac
     2401 ccccgaggtg acctgcgtgg tggtggccgt gtcccacgag gacccagagg tgaagttcaa
     2461 ctggtacgtg gacggcgtgg aggtgcacaa cgccaagacc aagcccagag aggagcagta
     2521 caacagcacc tacagggtgg tgtccgtgct gaccgtgctg caccaggact ggctgaacgg
     2581 caaagaatac aagtgcaaag tctccaacaa ggccctggct gccccaatcg aaaagacaat
     2641 cagcaaggcc aagggccagc cacgggagcc ccaggtgtac accctgcccc ccagccggga
     2701 ggagatgacc aagaaccagg tgtccctgac ctgtctggtg aagggcttct accccagcga
     2761 tatcgccgtg gagtgggaga gcaacggcca gcccgagaac aactacaaga ccaccccccc
     2821 agtgctggac agcgacggca gcttcttcct gtacagcaag ctgaccgtgg acaagtccag
     2881 gtggcagcag ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta
     2941 cacccagaag tccctgagcc tgagccccgg cggcagcggt cttaatgata ttttcgaggc
     3001 tcagaagatt gagtggcatg aacaccacca ccatcaccac taaggccggc ctgtagtcac
     3061 tggcacaacg cgttacccgg taagccaatc gggtatacac ggtcgtcata ctgcagacag
     3121 ggttcttcta ctttgcaaga tagtcttgag tagtaaaata aatagataga gaaaatcagc
     3181 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3241 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcaca
     3301 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3361 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3421 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3481 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3541 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3601 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3661 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3721 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3781 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     3841 cgacaaggtg aggaactaaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3901 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3961 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4021 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4081 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4141 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4201 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4261 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4321 cgccggggtg cccgccttcc tggagacatc cgcgccccgc aacctcccct tctacgagcg
     4381 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4441 gacccgcaag cccggtgcct gacacgtgct acgagatttc gattccaccg ccgccttcta
     4501 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4561 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4621 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4681 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4741 ctagagcttg gcgtaatcat ggtcattacc aatgcttaat cagtgaggca cctatctcag
     4801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4861 tacgggaggg cttaccatct ggccccagcg ctgcgatgat accgcgagaa ccacgctcac
     4921 cggctccgga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5041 gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac
     5101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5281 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatattc ttcctttttc
     5641 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5701 tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa ttaaccaatt ctgaacatta
     5761 tcgcgagccc atttatacct gaatatggct cataacaccc cttgctcatg accaaaatcc
     5821 cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5881 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5941 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6001 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagc
     6061 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6121 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6181 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6241 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6301 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6361 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6421 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6481 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6541 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6601 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg
     6661 cgagagtagg gaactgccag gcatcaaact aagcagaagg cccctgacgg atggcctttt
     6721 tgcgtttcta caaactcttt ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc
     6781 ctgggcggtt ctgataacga gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg
     6841 cgggtttttt tatgggggga gtttagggaa agagcatttg tcagaatatt taagggcgcc
     6901 tgtcactttg cttgatatat gagaattatt taaccttata aatgagaaaa aagcaacgca
     6961 ctttaaataa gatacgttgc tttttcgatt gatgaacacc tataattaaa ctattcatct
     7021 attatttatg attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat
     7081 aaatctcgaa aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg
     7141 ataatacaaa atataataca aactataaga tgttatcagt atttattatc atttagaata
     7201 aattttgtgt cgcccttaat tgtgagcgga taacaattac gagcttcatg cacagtggcg
     7261 ttgactcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
     7321 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     7381 tgtccaatat gaccgccatg tt
//
