LOCUS       sequence-368509-        6550 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 6550)
  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 6550)
  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-FAS-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6550
                     /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(1931..1950)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1963..1981)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1964..1981)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2191..2215)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2212..2415)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2241..2261)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2416..2795
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2962..2978
                     /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(3434..3450)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3549..3566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3720..4308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3800..3819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4394..4413)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4419..4438)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4480..4571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4572..4640,4641..5432)
                     /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(4790..4809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5485..5606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5496..5515)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5550..5569
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5736..6335)
                     /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(5819..5839)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6316..6335
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6354..6401)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6449..6550,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6463..6550,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6517..6536)
                     /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 cggcgcgccg ttggaccggc
     1381 tgccttcctc tttgcacttg gtattggagg tcagggtgca ctctttgatg atgccgtgtt
     1441 cgcactttgt gcaagggtcg cagtgctcgc acacggtgct gttgcagaag aagttgggct
     1501 tgcaccggca cttggtgttc tgtgtccggg tgcagttgat ttccacttcc aggccgtggc
     1561 cttcatcaca cagtctgcac cgtctgcact tgctgctgaa gtgggccttg tcggtgtact
     1621 ctttgccctc ttgacagggc acgcaatcgg gctcgtcgcc attcactgtg caatctctgg
     1681 ccttccgctc gccaggtgga caaggtttgt gacagaactg gccgtcgtgg tgcagtcctt
     1741 ccaggttctg ggtttccacg gtggtcacgg tctttctcag ttccaggcct ttgctattga
     1801 tgtcggtcac ttgggcggcc gcagcgaaca ccaggcagaa gatgtagctc agggcaatga
     1861 tggtcttcat ggtggcgcta gcaggcgcgt tttcgataga accgaggtgc agttggacct
     1921 gggagtggac acctgtggag agaaaggcaa agtggatgtt attctatagt gtcacctaaa
     1981 tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta acgaagccaa
     2041 gggggtgggc ctatagactc tataggcggt acttacgtca ctcttggcac ggggaatccg
     2101 cgttccaatg caccgttccc ggccgcggag gctggatcgg tcccggtgtc ttctatggag
     2161 gtcaaaacag cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctctgct
     2221 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     2281 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2341 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2401 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2461 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2521 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2581 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2641 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2701 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     2761 tgattactat taataactag tcaataatca atgccaacat ggcggtcata ttggacatga
     2821 gccaatataa atgtacatat tatgatatag atacaacgta tgcaatggcc aatagccaat
     2881 attgatttat gctatataac caatgaataa tatggctaat ggccaatatt gagtcaacgc
     2941 cactgtgcat gaagctcgta attgttatcc gctcacaatt aagggcgaca caaaatttat
     3001 tctaaatgat aataaatact gataacatct tatagtttgt attatatttt gtattatcgt
     3061 tgacatgtat aattttgata tcaaaaactg attttccctt tattattttc gagatttatt
     3121 ttcttaattc tctttaacaa actagaaata ttgtatatac aaaaaatcat aaataataga
     3181 tgaatagttt aattataggt gttcatcaat cgaaaaagca acgtatctta tttaaagtgc
     3241 gttgcttttt tctcatttat aaggttaaat aattctcata tatcaagcaa agtgacaggc
     3301 gcccttaaat attctgacaa atgctctttc cctaaactcc ccccataaaa aaacccgccg
     3361 aagcgggttt ttacgttatt tgcggattaa cgattactcg ttatcagaac cgcccagggg
     3421 gcccgagctt aagactggcc gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa
     3481 aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct actctcgcct
     3541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtgggct
     4141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4321 atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga
     4381 ttttggtcat gagcaagggg tgttatgagc catattcagg tataaatggg ctcgcgataa
     4441 tgttcagaat tggttaattg gttgtaacac tgacccctat ttgtttattt ttctaaatac
     4501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4561 aaaggaagaa tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4621 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4681 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4741 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4801 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4861 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4921 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4981 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5041 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5101 acaccacgat gcctgtagcg atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5161 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5221 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatcc ggagccggtg
     5281 agcgtggttc tcgcggtatc atcgcagcgc tggggccaga tggtaagccc tcccgtatcg
     5341 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5401 agataggtgc ctcactgatt aagcattggt aatgaccatg attacgccaa gctctagcta
     5461 gaggtcgacg gtatacagac atgataagat acattgatga gtttggacaa accacaacta
     5521 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     5581 ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg
     5641 cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag
     5701 aaggcggcgg tggaatcgaa atctcgtagc acgtgtcagg caccgggctt gcgggtcatg
     5761 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     5821 tcgtagaagg ggaggttgcg gggcgcggat gtctccagga aggcgggcac cccggcgcgc
     5881 tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
     5941 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     6001 aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     6061 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     6121 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
     6181 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
     6241 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
     6301 gtggcgaggc gcaccgtggg cttgtactcg gtcatggttt agttcctcac cttgtcgtat
     6361 tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga tcagatccga
     6421 aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca aaaaagcctc
     6481 ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca taaataaaaa
     6541 aaattagtca
//
