LOCUS       sequence-368602-        6517 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 6517)
  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 6517)
  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-RELT-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     202..222
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    340..451
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     440..457
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(597..614)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(615..659)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1887..1906)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1919..1937)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1920..1937)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2147..2171)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2168..2371)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2197..2217)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2372..2751
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2918..2934
                     /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(3390..3407)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3390..3406)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3505..3522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3676..4264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3756..3775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4350..4369)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4375..4394)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4436..4527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4528..4596,4597..5388)
                     /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(4746..4765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5441..5562
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5452..5471)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5506..5525
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5692..6291)
                     /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(5775..5795)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6272..6291
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6310..6357)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6405..6517,1..217))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6419..6517,1..37)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6473..6492)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
       61 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
      121 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      181 gggagcctgg ggactttcca caccctaact gacacacatt ccacagaatt aattcgcgtt
      241 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
      301 taaatcaaaa gaatagaccg agatagggtt gagtgttgtg atgcaatttc ctcattttat
      361 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
      421 acaacagatg gctggcaact agaaggcaca gtcgaggctg attttctcta tctatttatt
      481 ttactactca agactatctt gcaaagtaga agaaccctgt ctgcagtatg acgaccgtgt
      541 atacccgatt ggcttaccgg gtaacgcgtt gtgccagtga ctacaggccg gccttagtgg
      601 tgatggtggt ggtgttcatg ccactcaatc ttctgagcct cgaaaatatc attaagaccg
      661 ctgccgccgg ggctcaggct cagggacttc tgggtgtagt ggttgtgcag ggcctcgtgc
      721 atcacgctgc agctgaacac gttgccctgc tgccacctgg acttgtccac ggtcagcttg
      781 ctgtacagga agaagctgcc gtcgctgtcc agcactgggg gggtggtctt gtagttgttc
      841 tcgggctggc cgttgctctc ccactccacg gcgatatcgc tggggtagaa gcccttcacc
      901 agacaggtca gggacacctg gttcttggtc atctcctccc ggctgggggg cagggtgtac
      961 acctggggct cccgtggctg gcccttggcc ttgctgattg tcttttcgat tggggcagcc
     1021 agggccttgt tggagacttt gcacttgtat tctttgccgt tcagccagtc ctggtgcagc
     1081 acggtcagca cggacaccac cctgtaggtg ctgttgtact gctcctctct gggcttggtc
     1141 ttggcgttgt gcacctccac gccgtccacg taccagttga acttcacctc tgggtcctcg
     1201 tgggacacgg ccaccaccac gcaggtcacc tcgggggtcc tgctgatcat cagggtgtcc
     1261 ttgggcttgg gggggaacag gaacacggaa ggccctccca gcagttctgg agctgggcag
     1321 ggggggcagg tgtgggtctt gtcgccgccc ggcgcgcctt gagcagcggt ttcttcaggt
     1381 ccgccggctc ttgttccgtt gccaggctgt cttgtttcgc cgccagaaga agcgccagca
     1441 gcaacttcaa cgcctcttct agctctgcgg ccccattcat cacatccgtg tgttcccaga
     1501 ggggcccaag aacaaggctg gcaaggaact ctaggcactc cccaaggtcc gaaccatcca
     1561 ggccagcaat ctccgcaaag ggtatctctg gtggccatgc caacttgggc ttccagtctc
     1621 cgccacaggc tacatctggc atgaggctga caaggagaac tgccccaggc ggcagaaaat
     1681 gttccaggag gacaaggtct acacagggtg ccctgtccag gatcgagatc aggttcctcg
     1741 cctggaggac attgccacag tgttgtagag gcggccgcag cgaacaccag gcagaagatg
     1801 tagctcaggg caatgatggt cttcatggtg gcgctagcag gcgcgttttc gatagaaccg
     1861 aggtgcagtt ggacctggga gtggacacct gtggagagaa aggcaaagtg gatgttattc
     1921 tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc
     1981 gttctaacga agccaagggg gtgggcctat agactctata ggcggtactt acgtcactct
     2041 tggcacgggg aatccgcgtt ccaatgcacc gttcccggcc gcggaggctg gatcggtccc
     2101 ggtgtcttct atggaggtca aaacagcgtg gatggcgtct ccaggcgatc tgacggttca
     2161 ctaaacgagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     2221 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     2281 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     2341 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     2401 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     2461 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     2521 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     2581 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     2641 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     2701 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgc caacatggcg
     2761 gtcatattgg acatgagcca atataaatgt acatattatg atatagatac aacgtatgca
     2821 atggccaata gccaatattg atttatgcta tataaccaat gaataatatg gctaatggcc
     2881 aatattgagt caacgccact gtgcatgaag ctcgtaattg ttatccgctc acaattaagg
     2941 gcgacacaaa atttattcta aatgataata aatactgata acatcttata gtttgtatta
     3001 tattttgtat tatcgttgac atgtataatt ttgatatcaa aaactgattt tccctttatt
     3061 attttcgaga tttattttct taattctctt taacaaacta gaaatattgt atatacaaaa
     3121 aatcataaat aatagatgaa tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt
     3181 atcttattta aagtgcgttg cttttttctc atttataagg ttaaataatt ctcatatatc
     3241 aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc tctttcccta aactcccccc
     3301 ataaaaaaac ccgccgaagc gggtttttac gttatttgcg gattaacgat tactcgttat
     3361 cagaaccgcc cagggggccc gagcttaaga ctggccgtcg ttttacaaca cagaaagagt
     3421 ttgtagaaac gcaaaaaggc catccgtcag gggccttctg cttagtttga tgcctggcag
     3481 ttccctactc tcgccttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3541 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3601 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4081 cttgaagtgg tgggctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4201 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4261 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgcgcgt
     4321 aactcacgtt aagggatttt ggtcatgagc aaggggtgtt atgagccata ttcaggtata
     4381 aatgggctcg cgataatgtt cagaattggt taattggttg taacactgac ccctatttgt
     4441 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4501 cttcaataat attgaaaaag gaagaatatg agtattcaac atttccgtgt cgcccttatt
     4561 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4621 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4681 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4741 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4801 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4861 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4921 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4981 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5041 ccaaacgacg agcgtgacac cacgatgcct gtagcgatgg caacaacgtt gcgcaaacta
     5101 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5161 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5221 aaatccggag ccggtgagcg tggttctcgc ggtatcatcg cagcgctggg gccagatggt
     5281 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5341 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatg accatgatta
     5401 cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt
     5461 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     5521 attgctttat ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc
     5581 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     5641 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtagcacgt gtcaggcacc
     5701 gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac
     5761 ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggatgtct ccaggaaggc
     5821 gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc tgcccagacc
     5881 cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg
     5941 ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg aaccgctcaa
     6001 ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt
     6061 ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc
     6121 gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc
     6181 gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct
     6241 ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca tggtttagtt
     6301 cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat tgtcaacacg
     6361 tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca aaagcctagg
     6421 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc ggcctcggcc
     6481 tctgcataaa taaaaaaaat tagtcagcca tggggcg
//
