LOCUS       sequence-368510-        6553 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 6553)
  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 6553)
  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-CD8B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     170..190
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    308..419
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     408..425
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(565..582)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(583..627)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1891..1910)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1923..1941)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1924..1941)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2151..2175)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2172..2375)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2201..2221)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2376..2755
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2922..2938
                     /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(3394..3411)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3394..3410)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3509..3526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3680..4268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3760..3779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4354..4373)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4379..4398)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4440..4531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4532..4600,4601..5392)
                     /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(4750..4769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5445..5566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5456..5475)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5510..5529
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5696..6295)
                     /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(5779..5799)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6276..6295
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6314..6361)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6409..6553,1..185))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6423..6553,1..5)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6477..6496)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
       61 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
      121 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
      181 cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
      241 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
      301 gtgttgtgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag
      361 ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt
      421 cgaggctgat tttctctatc tatttatttt actactcaag actatcttgc aaagtagaag
      481 aaccctgtct gcagtatgac gaccgtgtat acccgattgg cttaccgggt aacgcgttgt
      541 gccagtgact acaggccggc cttagtggtg atggtggtgg tgttcatgcc actcaatctt
      601 ctgagcctcg aaaatatcat taagaccgct gccgccgggg ctcaggctca gggacttctg
      661 ggtgtagtgg ttgtgcaggg cctcgtgcat cacgctgcag ctgaacacgt tgccctgctg
      721 ccacctggac ttgtccacgg tcagcttgct gtacaggaag aagctgccgt cgctgtccag
      781 cactgggggg gtggtcttgt agttgttctc gggctggccg ttgctctccc actccacggc
      841 gatatcgctg gggtagaagc ccttcaccag acaggtcagg gacacctggt tcttggtcat
      901 ctcctcccgg ctggggggca gggtgtacac ctggggctcc cgtggctggc ccttggcctt
      961 gctgattgtc ttttcgattg gggcagccag ggccttgttg gagactttgc acttgtattc
     1021 tttgccgttc agccagtcct ggtgcagcac ggtcagcacg gacaccaccc tgtaggtgct
     1081 gttgtactgc tcctctctgg gcttggtctt ggcgttgtgc acctccacgc cgtccacgta
     1141 ccagttgaac ttcacctctg ggtcctcgtg ggacacggcc accaccacgc aggtcacctc
     1201 gggggtcctg ctgatcatca gggtgtcctt gggcttgggg gggaacagga acacggaagg
     1261 ccctcccagc agttctggag ctgggcaggg ggggcaggtg tgggtcttgt cgccgcccgg
     1321 cgcgccaggg ctacacagag ggcctttctg agtctcaggt ctgggcagtc tgcacactct
     1381 tttcttcagg gtagacttct tggtaggctg ggctgttgta ggcagaaagt ccaccacgct
     1441 cagctgtgtt cccttgccaa atgtcagctc agggctgccc acgatcatgc agaaatagat
     1501 gccgctgtcc tcgggcttca cgcttgtcag gttcaggatg aatctgctgg cgtcccggaa
     1561 cacggcgatc ttctcttgtt ccacttcctc gccgtgaatt gtgcccttgg cgctatccca
     1621 cagggccaga aattcatggt ggctatcgct gctaggggcc tgtctctgtc tcagccagta
     1681 gatccgcatg ttgctcaggc tgatcttggc ctcgcagctc agcatgacca tcttgttggt
     1741 ctgcaccttg atataggctg gtgtctgctg caaggcggcc gcagcgaaca ccaggcagaa
     1801 gatgtagctc agggcaatga tggtcttcat ggtggcgcta gcaggcgcgt tttcgataga
     1861 accgaggtgc agttggacct gggagtggac acctgtggag agaaaggcaa agtggatgtt
     1921 attctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag
     1981 ccgcgttcta acgaagccaa gggggtgggc ctatagactc tataggcggt acttacgtca
     2041 ctcttggcac ggggaatccg cgttccaatg caccgttccc ggccgcggag gctggatcgg
     2101 tcccggtgtc ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg
     2161 ttcactaaac gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     2221 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     2281 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     2341 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     2401 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     2461 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     2521 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     2581 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     2641 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     2701 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgccaacat
     2761 ggcggtcata ttggacatga gccaatataa atgtacatat tatgatatag atacaacgta
     2821 tgcaatggcc aatagccaat attgatttat gctatataac caatgaataa tatggctaat
     2881 ggccaatatt gagtcaacgc cactgtgcat gaagctcgta attgttatcc gctcacaatt
     2941 aagggcgaca caaaatttat tctaaatgat aataaatact gataacatct tatagtttgt
     3001 attatatttt gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt
     3061 tattattttc gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac
     3121 aaaaaatcat aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca
     3181 acgtatctta tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata
     3241 tatcaagcaa agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc
     3301 ccccataaaa aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg
     3361 ttatcagaac cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa
     3421 gagtttgtag aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg
     3481 gcagttccct actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3541 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3601 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4081 agttcttgaa gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg
     4141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc
     4321 gcgtaactca cgttaaggga ttttggtcat gagcaagggg tgttatgagc catattcagg
     4381 tataaatggg ctcgcgataa tgttcagaat tggttaattg gttgtaacac tgacccctat
     4441 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4501 aatgcttcaa taatattgaa aaaggaagaa tatgagtatt caacatttcc gtgtcgccct
     4561 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4621 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4681 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4741 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4801 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4861 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4921 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4981 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5041 cataccaaac gacgagcgtg acaccacgat gcctgtagcg atggcaacaa cgttgcgcaa
     5101 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5161 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5221 tgataaatcc ggagccggtg agcgtggttc tcgcggtatc atcgcagcgc tggggccaga
     5281 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5341 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aatgaccatg
     5401 attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga
     5461 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     5521 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga
     5581 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     5641 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtagc acgtgtcagg
     5701 caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg
     5761 tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggat gtctccagga
     5821 aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca
     5881 gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct
     5941 tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc
     6001 tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg
     6061 gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga
     6121 gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg
     6181 gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
     6241 ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggttt
     6301 agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
     6361 cacgtgctga tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc
     6421 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc
     6481 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     6541 cggagttagg ggc
//
