LOCUS       sequence-368515-        6565 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 6565)
  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 6565)
  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-CD8B2-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     97..116
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(135..182)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(230..559)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      244..379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(298..317)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     544..564
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    682..793
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     782..799
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(939..956)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(957..1001)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2277..2296)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2309..2327)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2310..2327)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2537..2561)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2558..2761)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2587..2607)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2762..3141
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3308..3324
                     /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(3780..3797)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3780..3796)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3895..3912)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4066..4654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4146..4165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4740..4759)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4765..4784)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4826..4917
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4918..4986,4987..5778)
                     /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(5136..5155)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5831..5952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5842..5861)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5896..5915
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6082..6565,1..116))
                     /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(6165..6185)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
       61 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt
      121 tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga ttaattgtca
      181 acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc
      241 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct
      301 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
      361 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
      421 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
      481 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      541 cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag
      601 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      661 cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag gacagtggga
      721 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
      781 ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact caagactatc
      841 ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga ttggcttacc
      901 gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg gtggtgttca
      961 tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc ggggctcagg
     1021 ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct gcagctgaac
     1081 acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag gaagaagctg
     1141 ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg gccgttgctc
     1201 tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt cagggacacc
     1261 tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg ctcccgtggc
     1321 tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt gttggagact
     1381 ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag cacggacacc
     1441 accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt gtgcacctcc
     1501 acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac ggccaccacc
     1561 acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt gggggggaac
     1621 aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca ggtgtgggtc
     1681 ttgtcgccgc ccggcgcgcc agggctacac agagggcctt tctgagtctc aggtctgggc
     1741 agtctgcaca ctcttttctt cagggtagac ttcttggtag gctgggctgt tgtaggcaga
     1801 aagtccacca ccacgctcag ctgtgttccc ttgccaaagg tcagctcagg gctgcccacg
     1861 atcatgcaga aatagatgcc gctgtcctcg ggcttcacgc ttgtcaggtt caggatgaat
     1921 ctgctggcgt cccggaacac ggcgatcttc tcttgttcca cttcctcgcc gtgaatggtg
     1981 cccttggcgc tatcccacag tgtcagaaac tcgtggtggc tatcgctgct aggggcctgt
     2041 ctctgtctca gccagtagat gcacatgttg ctcaggctga tcttggcctc gcagctcagc
     2101 atgaccatct tgttggtctg caccttgatg taggcagggg tctgctgcag aacagaattg
     2161 gcggccgcag cgaacaccag gcagaagatg tagctcaggg caatgatggt cttcatggtg
     2221 gcgctagcag gcgcgttttc gatagaaccg aggtgcagtt ggacctggga gtggacacct
     2281 gtggagagaa aggcaaagtg gatgttattc tatagtgtca cctaaatcgt atgtgtatga
     2341 tacataaggt tatgtattaa ttgtagccgc gttctaacga agccaagggg gtgggcctat
     2401 agactctata ggcggtactt acgtcactct tggcacgggg aatccgcgtt ccaatgcacc
     2461 gttcccggcc gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg
     2521 gatggcgtct ccaggcgatc tgacggttca ctaaacgagc tctgcttata tagacctccc
     2581 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     2641 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     2701 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     2761 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     2821 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     2881 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     2941 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     3001 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     3061 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     3121 aactagtcaa taatcaatgc caacatggcg gtcatattgg acatgagcca atataaatgt
     3181 acatattatg atatagatac aacgtatgca atggccaata gccaatattg atttatgcta
     3241 tataaccaat gaataatatg gctaatggcc aatattgagt caacgccact gtgcatgaag
     3301 ctcgtaattg ttatccgctc acaattaagg gcgacacaaa atttattcta aatgataata
     3361 aatactgata acatcttata gtttgtatta tattttgtat tatcgttgac atgtataatt
     3421 ttgatatcaa aaactgattt tccctttatt attttcgaga tttattttct taattctctt
     3481 taacaaacta gaaatattgt atatacaaaa aatcataaat aatagatgaa tagtttaatt
     3541 ataggtgttc atcaatcgaa aaagcaacgt atcttattta aagtgcgttg cttttttctc
     3601 atttataagg ttaaataatt ctcatatatc aagcaaagtg acaggcgccc ttaaatattc
     3661 tgacaaatgc tctttcccta aactcccccc ataaaaaaac ccgccgaagc gggtttttac
     3721 gttatttgcg gattaacgat tactcgttat cagaaccgcc cagggggccc gagcttaaga
     3781 ctggccgtcg ttttacaaca cagaaagagt ttgtagaaac gcaaaaaggc catccgtcag
     3841 gggccttctg cttagtttga tgcctggcag ttccctactc tcgccttccg cttcctcgct
     3901 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tgggctaact acggctacac
     4501 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4681 gtctgacgct cagtggaacg acgcgcgcgt aactcacgtt aagggatttt ggtcatgagc
     4741 aaggggtgtt atgagccata ttcaggtata aatgggctcg cgataatgtt cagaattggt
     4801 taattggttg taacactgac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4861 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatatg
     4921 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4981 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5041 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5101 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5161 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5221 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5281 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5341 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5401 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5461 gtagcgatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5521 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5581 gcccttccgg ctggctggtt tattgctgat aaatccggag ccggtgagcg tggttctcgc
     5641 ggtatcatcg cagcgctggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5701 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5761 ctgattaagc attggtaatg accatgatta cgccaagctc tagctagagg tcgacggtat
     5821 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     5881 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     5941 aataaacaag ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     6001 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     6061 atcgaaatct cgtagcacgt gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg
     6121 tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag
     6181 gttgcggggc gcggatgtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac
     6241 tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac
     6301 ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg
     6361 ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa
     6421 caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc
     6481 cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc
     6541 ggtgacccgc tcgatgtggc ggtcc
//
