LOCUS       sequence-368841-        6487 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 6487)
  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 6487)
  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-BTLA-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    351..462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     451..468
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(608..625)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(626..670)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1868..1887)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1900..1918)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1901..1918)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2128..2152)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2149..2352)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2178..2198)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2353..2732
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2899..2915
                     /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)."
     primer_bind     complement(3371..3388)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3371..3387)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3486..3503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3657..4245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3737..3756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4331..4350)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4356..4375)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4417..4508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4509..4577,4578..5369)
                     /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     complement(4727..4746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5422..5543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5433..5452)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5487..5506
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5673..6272)
                     /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(5756..5776)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6253..6272
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6291..6338)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6386..6487,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6400..6487,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6454..6473)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
      241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt gatgcaattt
      361 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
      421 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gattttctct
      481 atctatttat tttactactc aagactatct tgcaaagtag aagaaccctg tctgcagtat
      541 gacgaccgtg tatacccgat tggcttaccg ggtaacgcgt tgtgccagtg actacaggcc
      601 ggccttagtg gtgatggtgg tggtgttcat gccactcaat cttctgagcc tcgaaaatat
      661 cattaagacc gctgccgccg gggctcaggc tcagggactt ctgggtgtag tggttgtgca
      721 gggcctcgtg catcacgctg cagctgaaca cgttgccctg ctgccacctg gacttgtcca
      781 cggtcagctt gctgtacagg aagaagctgc cgtcgctgtc cagcactggg ggggtggtct
      841 tgtagttgtt ctcgggctgg ccgttgctct cccactccac ggcgatatcg ctggggtaga
      901 agcccttcac cagacaggtc agggacacct ggttcttggt catctcctcc cggctggggg
      961 gcagggtgta cacctggggc tcccgtggct ggcccttggc cttgctgatt gtcttttcga
     1021 ttggggcagc cagggccttg ttggagactt tgcacttgta ttctttgccg ttcagccagt
     1081 cctggtgcag cacggtcagc acggacacca ccctgtaggt gctgttgtac tgctcctctc
     1141 tgggcttggt cttggcgttg tgcacctcca cgccgtccac gtaccagttg aacttcacct
     1201 ctgggtcctc gtgggacacg gccaccacca cgcaggtcac ctcgggggtc ctgctgatca
     1261 tcagggtgtc cttgggcttg ggggggaaca ggaacacgga aggccctccc agcagttctg
     1321 gagctgggca gggggggcag gtgtgggtct tgtcgccgcc cggcgcgcct ctgtacagca
     1381 gccaaggccg ggaagccatc tcatccttag aaggcctctc gctggcggac ttcacatcgg
     1441 tcacgtacag tgtggtgctg tggctctcga tcaggttgct ctggaagttg gcgctgcatc
     1501 tgtagctgcc attgtcgttg ggcagcacgg gctcgaagtg caggatgaag aagctgatat
     1561 tcttttcctc tttccagctg gtctgccgat cttccagctt cacgcatgtg gtgccgttca
     1621 gcttgcacca ggtcacgtga ggtctgttgg cgcagtactt cacagggcat tccagctcga
     1681 aaggatcgcc ggccagaata gagtgctcgc tctgccgctt gatgtacagc tgcacgtcac
     1741 agctttcttt ggcggccgca gcgaacacca ggcagaagat gtagctcagg gcaatgatgg
     1801 tcttcatggt ggcgctagca ggcgcgtttt cgatagaacc gaggtgcagt tggacctggg
     1861 agtggacacc tgtggagaga aaggcaaagt ggatgttatt ctatagtgtc acctaaatcg
     1921 tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg aagccaaggg
     1981 ggtgggccta tagactctat aggcggtact tacgtcactc ttggcacggg gaatccgcgt
     2041 tccaatgcac cgttcccggc cgcggaggct ggatcggtcc cggtgtcttc tatggaggtc
     2101 aaaacagcgt ggatggcgtc tccaggcgat ctgacggttc actaaacgag ctctgcttat
     2161 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     2221 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     2281 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     2341 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     2401 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     2461 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     2521 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     2581 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     2641 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     2701 ttactattaa taactagtca ataatcaatg ccaacatggc ggtcatattg gacatgagcc
     2761 aatataaatg tacatattat gatatagata caacgtatgc aatggccaat agccaatatt
     2821 gatttatgct atataaccaa tgaataatat ggctaatggc caatattgag tcaacgccac
     2881 tgtgcatgaa gctcgtaatt gttatccgct cacaattaag ggcgacacaa aatttattct
     2941 aaatgataat aaatactgat aacatcttat agtttgtatt atattttgta ttatcgttga
     3001 catgtataat tttgatatca aaaactgatt ttccctttat tattttcgag atttattttc
     3061 ttaattctct ttaacaaact agaaatattg tatatacaaa aaatcataaa taatagatga
     3121 atagtttaat tataggtgtt catcaatcga aaaagcaacg tatcttattt aaagtgcgtt
     3181 gcttttttct catttataag gttaaataat tctcatatat caagcaaagt gacaggcgcc
     3241 cttaaatatt ctgacaaatg ctctttccct aaactccccc cataaaaaaa cccgccgaag
     3301 cgggttttta cgttatttgc ggattaacga ttactcgtta tcagaaccgc ccagggggcc
     3361 cgagcttaag actggccgtc gttttacaac acagaaagag tttgtagaaa cgcaaaaagg
     3421 ccatccgtca ggggccttct gcttagtttg atgcctggca gttccctact ctcgccttcc
     3481 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3541 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3601 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3661 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3721 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3781 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3841 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3901 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3961 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4021 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtgggctaac
     4081 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4141 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4201 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4261 ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt taagggattt
     4321 tggtcatgag caaggggtgt tatgagccat attcaggtat aaatgggctc gcgataatgt
     4381 tcagaattgg ttaattggtt gtaacactga cccctatttg tttatttttc taaatacatt
     4441 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4501 ggaagaatat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4561 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4621 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4681 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4741 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4801 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4861 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4921 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4981 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5041 ccacgatgcc tgtagcgatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5101 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5161 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatccgga gccggtgagc
     5221 gtggttctcg cggtatcatc gcagcgctgg ggccagatgg taagccctcc cgtatcgtag
     5281 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5341 taggtgcctc actgattaag cattggtaat gaccatgatt acgccaagct ctagctagag
     5401 gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5461 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     5521 ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc
     5581 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     5641 gcggcggtgg aatcgaaatc tcgtagcacg tgtcaggcac cgggcttgcg ggtcatgcac
     5701 caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg
     5761 tagaagggga ggttgcgggg cgcggatgtc tccaggaagg cgggcacccc ggcgcgctcg
     5821 gccgcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc
     5881 gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg
     5941 ccttccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg
     6001 atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg
     6061 gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt
     6121 tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg
     6181 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg
     6241 gcgaggcgca ccgtgggctt gtactcggtc atggtttagt tcctcacctt gtcgtattat
     6301 actatgccga tatactatgc cgatgattaa ttgtcaacac gtgctgatca gatccgaaaa
     6361 tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa aagcctcctc
     6421 actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa
     6481 ttagtca
//
