LOCUS       sequence-368836-        6484 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 6484)
  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 6484)
  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-IGSF6-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..216
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    334..445
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     434..451
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(591..608)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(609..653)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1848..1867)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1880..1898)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1881..1898)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2108..2132)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2129..2332)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2158..2178)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2333..2712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2879..2895
                     /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(3351..3368)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3351..3367)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3466..3483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3637..4225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3717..3736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4311..4330)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4336..4355)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4397..4488
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4489..4557,4558..5349)
                     /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(4707..4726)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5402..5523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5413..5432)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5467..5486
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5653..6252)
                     /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(5736..5756)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6233..6252
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6271..6318)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6366..6484,1..211))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6380..6484,1..31)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6434..6453)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
       61 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      121 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      181 ctggggactt tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt
      241 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      301 aaaagaatag accgagatag ggttgagtgt tgtgatgcaa tttcctcatt ttattaggaa
      361 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
      421 gatggctggc aactagaagg cacagtcgag gctgattttc tctatctatt tattttacta
      481 ctcaagacta tcttgcaaag tagaagaacc ctgtctgcag tatgacgacc gtgtataccc
      541 gattggctta ccgggtaacg cgttgtgcca gtgactacag gccggcctta gtggtgatgg
      601 tggtggtgtt catgccactc aatcttctga gcctcgaaaa tatcattaag accgctgccg
      661 ccggggctca ggctcaggga cttctgggtg tagtggttgt gcagggcctc gtgcatcacg
      721 ctgcagctga acacgttgcc ctgctgccac ctggacttgt ccacggtcag cttgctgtac
      781 aggaagaagc tgccgtcgct gtccagcact gggggggtgg tcttgtagtt gttctcgggc
      841 tggccgttgc tctcccactc cacggcgata tcgctggggt agaagccctt caccagacag
      901 gtcagggaca cctggttctt ggtcatctcc tcccggctgg ggggcagggt gtacacctgg
      961 ggctcccgtg gctggccctt ggccttgctg attgtctttt cgattggggc agccagggcc
     1021 ttgttggaga ctttgcactt gtattctttg ccgttcagcc agtcctggtg cagcacggtc
     1081 agcacggaca ccaccctgta ggtgctgttg tactgctcct ctctgggctt ggtcttggcg
     1141 ttgtgcacct ccacgccgtc cacgtaccag ttgaacttca cctctgggtc ctcgtgggac
     1201 acggccacca ccacgcaggt cacctcgggg gtcctgctga tcatcagggt gtccttgggc
     1261 ttggggggga acaggaacac ggaaggccct cccagcagtt ctggagctgg gcaggggggg
     1321 caggtgtggg tcttgtcgcc gcccggcgcg ccagatctca gctctttgct cagcagcttg
     1381 atttcccgca cgaccagtgt tgttccgccg cctgtctgtt tagctctggc ctctggcacg
     1441 cttggaaagg cgattccgca gatgtagatg gcgctgtcgt tgctggtcac tctgttcacg
     1501 gtcagggaca cctggttttc tttcagggcc tcgcgcactg taaacttatc ggcctcgctc
     1561 ttgcagccat ccaggcacag attctcaggc tggtgggcgc catatctgaa ccacaggcag
     1621 gtaggttgct cgctagggca gcctgtggcg ctgaatgtac acttgatggt cacggcctcg
     1681 tgggtgtagt ccacttccag ataccaaggc tgtgtcacag acagtgtaca ggcggccgca
     1741 gcgaacacca ggcagaagat gtagctcagg gcaatgatgg tcttcatggt ggcgctagca
     1801 ggcgcgtttt cgatagaacc gaggtgcagt tggacctggg agtggacacc tgtggagaga
     1861 aaggcaaagt ggatgttatt ctatagtgtc acctaaatcg tatgtgtatg atacataagg
     1921 ttatgtatta attgtagccg cgttctaacg aagccaaggg ggtgggccta tagactctat
     1981 aggcggtact tacgtcactc ttggcacggg gaatccgcgt tccaatgcac cgttcccggc
     2041 cgcggaggct ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt ggatggcgtc
     2101 tccaggcgat ctgacggttc actaaacgag ctctgcttat atagacctcc caccgtacac
     2161 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     2221 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     2281 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     2341 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     2401 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     2461 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     2521 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     2581 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     2641 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     2701 ataatcaatg ccaacatggc ggtcatattg gacatgagcc aatataaatg tacatattat
     2761 gatatagata caacgtatgc aatggccaat agccaatatt gatttatgct atataaccaa
     2821 tgaataatat ggctaatggc caatattgag tcaacgccac tgtgcatgaa gctcgtaatt
     2881 gttatccgct cacaattaag ggcgacacaa aatttattct aaatgataat aaatactgat
     2941 aacatcttat agtttgtatt atattttgta ttatcgttga catgtataat tttgatatca
     3001 aaaactgatt ttccctttat tattttcgag atttattttc ttaattctct ttaacaaact
     3061 agaaatattg tatatacaaa aaatcataaa taatagatga atagtttaat tataggtgtt
     3121 catcaatcga aaaagcaacg tatcttattt aaagtgcgtt gcttttttct catttataag
     3181 gttaaataat tctcatatat caagcaaagt gacaggcgcc cttaaatatt ctgacaaatg
     3241 ctctttccct aaactccccc cataaaaaaa cccgccgaag cgggttttta cgttatttgc
     3301 ggattaacga ttactcgtta tcagaaccgc ccagggggcc cgagcttaag actggccgtc
     3361 gttttacaac acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca ggggccttct
     3421 gcttagtttg atgcctggca gttccctact ctcgccttcc gcttcctcgc tcactgactc
     3481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4021 tgtaggcggt gctacagagt tcttgaagtg gtgggctaac tacggctaca ctagaagaac
     4081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4261 tcagtggaac gacgcgcgcg taactcacgt taagggattt tggtcatgag caaggggtgt
     4321 tatgagccat attcaggtat aaatgggctc gcgataatgt tcagaattgg ttaattggtt
     4381 gtaacactga cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4441 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatat gagtattcaa
     4501 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4561 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4621 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4681 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4741 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4801 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4861 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4921 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4981 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcgatg
     5041 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5101 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5161 gctggctggt ttattgctga taaatccgga gccggtgagc gtggttctcg cggtatcatc
     5221 gcagcgctgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5281 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5341 cattggtaat gaccatgatt acgccaagct ctagctagag gtcgacggta tacagacatg
     5401 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     5461 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     5521 gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     5581 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     5641 tcgtagcacg tgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg
     5701 cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg
     5761 cgcggatgtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag
     5821 cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag
     5881 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
     5941 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
     6001 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
     6061 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
     6121 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
     6181 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
     6241 gtactcggtc atggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc
     6301 cgatgattaa ttgtcaacac gtgctgatca gatccgaaaa tggatataca agctcccggg
     6361 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     6421 gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     6481 gaat
//
