LOCUS       sequence-368815-        6460 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 6460)
  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 6460)
  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-SIGIRR-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6460
                     /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(1851..1870)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1883..1901)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1884..1901)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2111..2135)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2132..2335)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2161..2181)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2336..2715
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2882..2898
                     /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(3354..3371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3354..3370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3469..3486)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3640..4228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3720..3739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4314..4333)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4339..4358)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4400..4491
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4492..4560,4561..5352)
                     /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(4710..4729)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5405..5526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5416..5435)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5470..5489
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5656..6255)
                     /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(5739..5759)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6236..6255
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6274..6321)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6369..6460,1..238))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6383..6460,1..58)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6437..6456)
                     /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 tggcttgtgg ggccagctct ctgcagtgtg aagctgctga agctgatgtt ctggatgcta
     1441 caggtgaagg cgccgtacac ctctgtgctg gtcacattca ctcccagcac gctagacacc
     1501 agcacctcgc tcagattggc tttgacccag ctgtactcgt gcaggctata gtgtccgccg
     1561 atgccgagag gcagtccatc cttcagccac tgcacgcttg gcagagaaca gtgagggcca
     1621 gacacaaccc aggctgtgca attcagggcc acagagcttc ccagagcagg tctgagcacc
     1681 tgatcctcag aggggctcag aaagtcaggg gctctgtcac aaacaccagg catggcggcc
     1741 gcagcgaaca ccaggcagaa gatgtagctc agggcaatga tggtcttcat ggtggcgcta
     1801 gcaggcgcgt tttcgataga accgaggtgc agttggacct gggagtggac acctgtggag
     1861 agaaaggcaa agtggatgtt attctatagt gtcacctaaa tcgtatgtgt atgatacata
     1921 aggttatgta ttaattgtag ccgcgttcta acgaagccaa gggggtgggc ctatagactc
     1981 tataggcggt acttacgtca ctcttggcac ggggaatccg cgttccaatg caccgttccc
     2041 ggccgcggag gctggatcgg tcccggtgtc ttctatggag gtcaaaacag cgtggatggc
     2101 gtctccaggc gatctgacgg ttcactaaac gagctctgct tatatagacc tcccaccgta
     2161 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     2221 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     2281 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     2341 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     2401 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     2461 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     2521 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     2581 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     2641 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     2701 tcaataatca atgccaacat ggcggtcata ttggacatga gccaatataa atgtacatat
     2761 tatgatatag atacaacgta tgcaatggcc aatagccaat attgatttat gctatataac
     2821 caatgaataa tatggctaat ggccaatatt gagtcaacgc cactgtgcat gaagctcgta
     2881 attgttatcc gctcacaatt aagggcgaca caaaatttat tctaaatgat aataaatact
     2941 gataacatct tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata
     3001 tcaaaaactg attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa
     3061 actagaaata ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt
     3121 gttcatcaat cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat
     3181 aaggttaaat aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa
     3241 atgctctttc cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt
     3301 tgcggattaa cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc
     3361 gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct
     3421 tctgcttagt ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga
     3481 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3541 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4021 gtatgtaggc ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag
     4081 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4201 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4261 cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcaagggg
     4321 tgttatgagc catattcagg tataaatggg ctcgcgataa tgttcagaat tggttaattg
     4381 gttgtaacac tgacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4441 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa tatgagtatt
     4501 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4561 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4621 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4681 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4741 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4801 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4861 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4921 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4981 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagcg
     5041 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5101 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5161 ccggctggct ggtttattgc tgataaatcc ggagccggtg agcgtggttc tcgcggtatc
     5221 atcgcagcgc tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5281 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5341 aagcattggt aatgaccatg attacgccaa gctctagcta gaggtcgacg gtatacagac
     5401 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     5461 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     5521 caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     5581 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     5641 atctcgtagc acgtgtcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc
     5701 gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg
     5761 gggcgcggat gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg
     5821 gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc
     5881 caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg
     5941 cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc
     6001 ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac
     6061 ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac
     6121 ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc
     6181 ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg
     6241 cttgtactcg gtcatggttt agttcctcac cttgtcgtat tatactatgc cgatatacta
     6301 tgccgatgat taattgtcaa cacgtgctga tcagatccga aaatggatat acaagctccc
     6361 gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     6421 tcagaggcag aggcggcctc ggcctctgca taaataaaaa
//
