LOCUS       sequence-368529-        6592 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 6592)
  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 6592)
  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-VSIR-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(145..192)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(240..569)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      254..389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(308..327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     554..574
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    692..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     792..809
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(949..966)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(967..1011)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2314..2333)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2346..2364)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2347..2364)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2574..2598)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2595..2798)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2624..2644)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2799..3178
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3345..3361
                     /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(3817..3834)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3817..3833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3932..3949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4103..4691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4183..4202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4777..4796)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4802..4821)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4863..4954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4955..5023,5024..5815)
                     /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(5173..5192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5868..5989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5879..5898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5933..5952
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6119..6592,1..126))
                     /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(6202..6222)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
       61 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
      121 ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
      181 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
      241 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
      301 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      361 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      421 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      481 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      541 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
      601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      661 aagaatagac cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag
      721 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      781 tggctggcaa ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact
      841 caagactatc ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga
      901 ttggcttacc gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg
      961 gtggtgttca tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc
     1021 ggggctcagg ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct
     1081 gcagctgaac acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag
     1141 gaagaagctg ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg
     1201 gccgttgctc tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt
     1261 cagggacacc tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg
     1321 ctcccgtggc tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt
     1381 gttggagact ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag
     1441 cacggacacc accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt
     1501 gtgcacctcc acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac
     1561 ggccaccacc acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt
     1621 gggggggaac aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca
     1681 ggtgtgggtc ttgtcgccgc ccggcgcgcc agcggctgtg atattctcgc tgtcctggct
     1741 gctgctgggg tacaccacac aattgctagg ggcgtccttg cctgtctgga cctgcagttc
     1801 catagcgccg tgcactctgt gttcgctgtg gtggtgtctg atttccacca ccagacagca
     1861 gtacaggccg ctatccagca gggtcagatt ccgcatggtg atgctgaagt tgccgtggtg
     1921 atcagaggca gattccaggc cgtgtctctg ggccagatca tgagaggtat tggcggcctg
     1981 atgtccgcca tgatgcagat gcaggtcctg gaaggtcagg ttccggatag gccttctctc
     2041 gctacaggtc tgcacctcgc ctctgctgga tctgtaccag gtcttgtaga aggtcacgtc
     2101 gtggccctta tccacggggc ccagcagtct acatgtcagg gtcacattct ggccctcagg
     2161 gcacacgtac aggctgtaag gtgtggccac cttaaaggcg gccgcagcga acaccaggca
     2221 gaagatgtag ctcagggcaa tgatggtctt catggtggcg ctagcaggcg cgttttcgat
     2281 agaaccgagg tgcagttgga cctgggagtg gacacctgtg gagagaaagg caaagtggat
     2341 gttattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
     2401 tagccgcgtt ctaacgaagc caagggggtg ggcctataga ctctataggc ggtacttacg
     2461 tcactcttgg cacggggaat ccgcgttcca atgcaccgtt cccggccgcg gaggctggat
     2521 cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga
     2581 cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     2641 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
     2701 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     2761 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     2821 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     2881 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     2941 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     3001 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     3061 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     3121 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgccaa
     3181 catggcggtc atattggaca tgagccaata taaatgtaca tattatgata tagatacaac
     3241 gtatgcaatg gccaatagcc aatattgatt tatgctatat aaccaatgaa taatatggct
     3301 aatggccaat attgagtcaa cgccactgtg catgaagctc gtaattgtta tccgctcaca
     3361 attaagggcg acacaaaatt tattctaaat gataataaat actgataaca tcttatagtt
     3421 tgtattatat tttgtattat cgttgacatg tataattttg atatcaaaaa ctgattttcc
     3481 ctttattatt ttcgagattt attttcttaa ttctctttaa caaactagaa atattgtata
     3541 tacaaaaaat cataaataat agatgaatag tttaattata ggtgttcatc aatcgaaaaa
     3601 gcaacgtatc ttatttaaag tgcgttgctt ttttctcatt tataaggtta aataattctc
     3661 atatatcaag caaagtgaca ggcgccctta aatattctga caaatgctct ttccctaaac
     3721 tccccccata aaaaaacccg ccgaagcggg tttttacgtt atttgcggat taacgattac
     3781 tcgttatcag aaccgcccag ggggcccgag cttaagactg gccgtcgttt tacaacacag
     3841 aaagagtttg tagaaacgca aaaaggccat ccgtcagggg ccttctgctt agtttgatgc
     3901 ctggcagttc cctactctcg ccttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4501 cagagttctt gaagtggtgg gctaactacg gctacactag aagaacagta tttggtatct
     4561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgacg
     4741 cgcgcgtaac tcacgttaag ggattttggt catgagcaag gggtgttatg agccatattc
     4801 aggtataaat gggctcgcga taatgttcag aattggttaa ttggttgtaa cactgacccc
     4861 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4921 ataaatgctt caataatatt gaaaaaggaa gaatatgagt attcaacatt tccgtgtcgc
     4981 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5041 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5101 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5161 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5221 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5281 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5341 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5401 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5461 agccatacca aacgacgagc gtgacaccac gatgcctgta gcgatggcaa caacgttgcg
     5521 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5581 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5641 tgctgataaa tccggagccg gtgagcgtgg ttctcgcggt atcatcgcag cgctggggcc
     5701 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5761 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaatgacc
     5821 atgattacgc caagctctag ctagaggtcg acggtataca gacatgataa gatacattga
     5881 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5941 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga
     6001 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     6061 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc
     6121 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     6181 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gatgtctcca
     6241 ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc
     6301 ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct
     6361 ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac
     6421 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
     6481 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
     6541 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg at
//
