LOCUS       sequence-368751-        6646 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 6646)
  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 6646)
  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-FCER1A-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6646
                     /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(2027..2046)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2059..2077)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2060..2077)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2287..2311)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2308..2511)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2337..2357)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2512..2891
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3058..3074
                     /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(3530..3547)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3530..3546)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3645..3662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3816..4404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3896..3915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4490..4509)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4515..4534)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4576..4667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4668..4736,4737..5528)
                     /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(4886..4905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5581..5702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5592..5611)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5646..5665
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5832..6431)
                     /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(5915..5935)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6412..6431
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6450..6497)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6545..6646,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6559..6646,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6613..6632)
                     /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 cggcgcgcct tgcagccagt
     1381 acttctctct aggggccttg atcacggtga tgttcagagg ctcggactcg tagtccagct
     1441 gccacacttt gccggtacag tagtaggtgc cgctatcttc cacggtggcg ttggtgatgc
     1501 tgatgttgtg gttctcgtac caatacttca gggcctcgcc gtccttgtag tagatcactt
     1561 tgtacacgtc ccagttccgc cagccgtggc atctcaggaa cagaggctgt ccttccatca
     1621 caacctcggc agaggcctgc agcagcagcc aatcagagaa cacttccagg tacacgggct
     1681 cgctctcgtt cacttgctgg tgctgacact tgtactcgcc gctgtcctcg aacttggcgt
     1741 tcacgatgtt caggctgctg tttgtttcct cgctcaggct gccgttgtgg aaccacttgg
     1801 tgctggacac ctcgaagaag ttgttgccgt tgcaggtcag ggtcacattc tcgcccttga
     1861 agatccggtt ccaaggaggg ttcagagaca ctttgggttt ctgaggaacg gcggccgcag
     1921 cgaacaccag gcagaagatg tagctcaggg caatgatggt cttcatggtg gcgctagcag
     1981 gcgcgttttc gatagaaccg aggtgcagtt ggacctggga gtggacacct gtggagagaa
     2041 aggcaaagtg gatgttattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt
     2101 tatgtattaa ttgtagccgc gttctaacga agccaagggg gtgggcctat agactctata
     2161 ggcggtactt acgtcactct tggcacgggg aatccgcgtt ccaatgcacc gttcccggcc
     2221 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
     2281 ccaggcgatc tgacggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     2341 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2401 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2461 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2521 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2581 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2641 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2701 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2761 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     2821 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa
     2881 taatcaatgc caacatggcg gtcatattgg acatgagcca atataaatgt acatattatg
     2941 atatagatac aacgtatgca atggccaata gccaatattg atttatgcta tataaccaat
     3001 gaataatatg gctaatggcc aatattgagt caacgccact gtgcatgaag ctcgtaattg
     3061 ttatccgctc acaattaagg gcgacacaaa atttattcta aatgataata aatactgata
     3121 acatcttata gtttgtatta tattttgtat tatcgttgac atgtataatt ttgatatcaa
     3181 aaactgattt tccctttatt attttcgaga tttattttct taattctctt taacaaacta
     3241 gaaatattgt atatacaaaa aatcataaat aatagatgaa tagtttaatt ataggtgttc
     3301 atcaatcgaa aaagcaacgt atcttattta aagtgcgttg cttttttctc atttataagg
     3361 ttaaataatt ctcatatatc aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc
     3421 tctttcccta aactcccccc ataaaaaaac ccgccgaagc gggtttttac gttatttgcg
     3481 gattaacgat tactcgttat cagaaccgcc cagggggccc gagcttaaga ctggccgtcg
     3541 ttttacaaca cagaaagagt ttgtagaaac gcaaaaaggc catccgtcag gggccttctg
     3601 cttagtttga tgcctggcag ttccctactc tcgccttccg cttcctcgct cactgactcg
     3661 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3721 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3781 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3841 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3901 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3961 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4021 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4081 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4141 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4201 gtaggcggtg ctacagagtt cttgaagtgg tgggctaact acggctacac tagaagaaca
     4261 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4321 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4381 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4441 cagtggaacg acgcgcgcgt aactcacgtt aagggatttt ggtcatgagc aaggggtgtt
     4501 atgagccata ttcaggtata aatgggctcg cgataatgtt cagaattggt taattggttg
     4561 taacactgac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatatg agtattcaac
     4681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5041 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcgatgg
     5221 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5281 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5341 ctggctggtt tattgctgat aaatccggag ccggtgagcg tggttctcgc ggtatcatcg
     5401 cagcgctggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5521 attggtaatg accatgatta cgccaagctc tagctagagg tcgacggtat acagacatga
     5581 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     5641 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5701 ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     5761 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     5821 cgtagcacgt gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     5881 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     5941 gcggatgtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     6001 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     6061 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     6121 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     6181 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     6241 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     6301 tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     6361 gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     6421 tactcggtca tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc
     6481 gatgattaat tgtcaacacg tgctgatcag atccgaaaat ggatatacaa gctcccggga
     6541 gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     6601 aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat tagtca
//
