LOCUS       sequence-368856-        6502 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 6502)
  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 6502)
  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-SCN4B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     223..243
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    361..472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     461..478
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(618..635)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(636..680)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1893..1912)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1925..1943)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1926..1943)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2153..2177)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2174..2377)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2203..2223)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2378..2757
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2924..2940
                     /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(3396..3413)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3396..3412)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3511..3528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3682..4270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3762..3781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4356..4375)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4381..4400)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4442..4533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4534..4602,4603..5394)
                     /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(4752..4771)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5447..5568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5458..5477)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5512..5531
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5698..6297)
                     /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(5781..5801)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6278..6297
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6316..6363)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6411..6502,1..238))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6425..6502,1..58)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6479..6498)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
       61 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
      121 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
      181 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
      241 tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
      301 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
      361 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
      421 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
      481 gattttctct atctatttat tttactactc aagactatct tgcaaagtag aagaaccctg
      541 tctgcagtat gacgaccgtg tatacccgat tggcttaccg ggtaacgcgt tgtgccagtg
      601 actacaggcc ggccttagtg gtgatggtgg tggtgttcat gccactcaat cttctgagcc
      661 tcgaaaatat cattaagacc gctgccgccg gggctcaggc tcagggactt ctgggtgtag
      721 tggttgtgca gggcctcgtg catcacgctg cagctgaaca cgttgccctg ctgccacctg
      781 gacttgtcca cggtcagctt gctgtacagg aagaagctgc cgtcgctgtc cagcactggg
      841 ggggtggtct tgtagttgtt ctcgggctgg ccgttgctct cccactccac ggcgatatcg
      901 ctggggtaga agcccttcac cagacaggtc agggacacct ggttcttggt catctcctcc
      961 cggctggggg gcagggtgta cacctggggc tcccgtggct ggcccttggc cttgctgatt
     1021 gtcttttcga ttggggcagc cagggccttg ttggagactt tgcacttgta ttctttgccg
     1081 ttcagccagt cctggtgcag cacggtcagc acggacacca ccctgtaggt gctgttgtac
     1141 tgctcctctc tgggcttggt cttggcgttg tgcacctcca cgccgtccac gtaccagttg
     1201 aacttcacct ctgggtcctc gtgggacacg gccaccacca cgcaggtcac ctcgggggtc
     1261 ctgctgatca tcagggtgtc cttgggcttg ggggggaaca ggaacacgga aggccctccc
     1321 agcagttctg gagctgggca gggggggcag gtgtgggtct tgtcgccgcc cggcgcgcca
     1381 acggtgttgt ccacctcttc cagtctgtcc accacttgca gaaagatggt ggcgtggtgc
     1441 tgcaggttgt tctctttggg attcttcacg tgacaggtgt acttgccggt gtcgctgaat
     1501 tccaggtcgc gcagcacgat gctgatgttg ttcatctttt ctttggtgct gcccacgagg
     1561 gtgattctgt cgtcgtcctt cagggtcact ttggggtcag acttctcgtt cttcacggtg
     1621 ccctcgatca ggatcttgaa ggcgtcgctg ctgttgtagg tccatctgaa gtgcaggtcc
     1681 tcgaagccga agcagctgga gaaggtacaa ggcagcagga tctcggtgcc gttcacggcg
     1741 tagatgtctg tggctttgcc cacagacact tccaaggcgg ccgcagcgaa caccaggcag
     1801 aagatgtagc tcagggcaat gatggtcttc atggtggcgc tagcaggcgc gttttcgata
     1861 gaaccgaggt gcagttggac ctgggagtgg acacctgtgg agagaaaggc aaagtggatg
     1921 ttattctata gtgtcaccta aatcgtatgt gtatgataca taaggttatg tattaattgt
     1981 agccgcgttc taacgaagcc aagggggtgg gcctatagac tctataggcg gtacttacgt
     2041 cactcttggc acggggaatc cgcgttccaa tgcaccgttc ccggccgcgg aggctggatc
     2101 ggtcccggtg tcttctatgg aggtcaaaac agcgtggatg gcgtctccag gcgatctgac
     2161 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     2221 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     2281 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     2341 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     2401 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     2461 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     2521 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     2581 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     2641 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     2701 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgccaac
     2761 atggcggtca tattggacat gagccaatat aaatgtacat attatgatat agatacaacg
     2821 tatgcaatgg ccaatagcca atattgattt atgctatata accaatgaat aatatggcta
     2881 atggccaata ttgagtcaac gccactgtgc atgaagctcg taattgttat ccgctcacaa
     2941 ttaagggcga cacaaaattt attctaaatg ataataaata ctgataacat cttatagttt
     3001 gtattatatt ttgtattatc gttgacatgt ataattttga tatcaaaaac tgattttccc
     3061 tttattattt tcgagattta ttttcttaat tctctttaac aaactagaaa tattgtatat
     3121 acaaaaaatc ataaataata gatgaatagt ttaattatag gtgttcatca atcgaaaaag
     3181 caacgtatct tatttaaagt gcgttgcttt tttctcattt ataaggttaa ataattctca
     3241 tatatcaagc aaagtgacag gcgcccttaa atattctgac aaatgctctt tccctaaact
     3301 ccccccataa aaaaacccgc cgaagcgggt ttttacgtta tttgcggatt aacgattact
     3361 cgttatcaga accgcccagg gggcccgagc ttaagactgg ccgtcgtttt acaacacaga
     3421 aagagtttgt agaaacgcaa aaaggccatc cgtcaggggc cttctgctta gtttgatgcc
     3481 tggcagttcc ctactctcgc cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4081 agagttcttg aagtggtggg ctaactacgg ctacactaga agaacagtat ttggtatctg
     4141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgacgc
     4321 gcgcgtaact cacgttaagg gattttggtc atgagcaagg ggtgttatga gccatattca
     4381 ggtataaatg ggctcgcgat aatgttcaga attggttaat tggttgtaac actgacccct
     4441 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4501 taaatgcttc aataatattg aaaaaggaag aatatgagta ttcaacattt ccgtgtcgcc
     4561 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4621 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4681 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4741 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4801 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4861 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4921 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4981 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5041 gccataccaa acgacgagcg tgacaccacg atgcctgtag cgatggcaac aacgttgcgc
     5101 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5161 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5221 gctgataaat ccggagccgg tgagcgtggt tctcgcggta tcatcgcagc gctggggcca
     5281 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5341 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaatgacca
     5401 tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag atacattgat
     5461 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     5521 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg ggtgggcgaa
     5581 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     5641 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgta gcacgtgtca
     5701 ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc
     5761 ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg atgtctccag
     5821 gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc
     5881 cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc
     5941 cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc
     6001 gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc
     6061 cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc
     6121 gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc
     6181 cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac
     6241 ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt
     6301 ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc
     6361 aacacgtgct gatcagatcc gaaaatggat atacaagctc ccgggagctt tttgcaaaag
     6421 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc agaggcggcc
     6481 tcggcctctg cataaataaa aa
//
