LOCUS       sequence-368524-        6574 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 6574)
  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 6574)
  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-CD300E-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(145..192)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(240..569)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      254..389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(308..327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     554..574
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    692..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     792..809
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(949..966)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(967..1011)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2296..2315)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2328..2346)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2329..2346)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2556..2580)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2577..2780)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2606..2626)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2781..3160
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3327..3343
                     /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(3799..3816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3799..3815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3914..3931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4165..4184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4759..4778)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4784..4803)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4845..4936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4937..5005,5006..5797)
                     /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(5155..5174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5850..5971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5861..5880)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5915..5934
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6101..6574,1..126))
                     /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(6184..6204)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
       61 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
      121 ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
      181 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
      241 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
      301 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      361 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      421 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      481 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      541 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
      601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      661 aagaatagac cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag
      721 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      781 tggctggcaa ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact
      841 caagactatc ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga
      901 ttggcttacc gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg
      961 gtggtgttca tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc
     1021 ggggctcagg ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct
     1081 gcagctgaac acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag
     1141 gaagaagctg ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg
     1201 gccgttgctc tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt
     1261 cagggacacc tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg
     1321 ctcccgtggc tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt
     1381 gttggagact ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag
     1441 cacggacacc accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt
     1501 gtgcacctcc acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac
     1561 ggccaccacc acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt
     1621 gggggggaac aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca
     1681 ggtgtgggtc ttgtcgccgc ccggcgcgcc atgagggctg ctcagtctaa agccgctgtt
     1741 ctgggtcagc acctcgcctg tgctcagatt tctgccgggg ttgaccacca gaaagattgg
     1801 aggtgtggcg gggtgtgttg tccgtctagg tgtggtgatg gcaggggaca cgtacacccg
     1861 cacgagatca gaaggatctc tggaccagct gtcgagcacc cacactgtct ggatcttgca
     1921 ccaatagctg ccggcatcgt cctcattcag gttctgcatg gtcacggtga aggccagagc
     1981 ctcaggatga tctctaatgg acacccggcc gttccgttcc accttctctt cgcccttagt
     2041 ttccacgatg ctctcgcagc tggtgtcgta ctggcctctg caccagtact tgttgtagcc
     2101 cttgtacatg ctctcgtact gacaccacac ggtcaggcta tcgccggctg tgccagtaac
     2161 agatccaggt cctttcaagg cggccgcagc gaacaccagg cagaagatgt agctcagggc
     2221 aatgatggtc ttcatggtgg cgctagcagg cgcgttttcg atagaaccga ggtgcagttg
     2281 gacctgggag tggacacctg tggagagaaa ggcaaagtgg atgttattct atagtgtcac
     2341 ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg ttctaacgaa
     2401 gccaaggggg tgggcctata gactctatag gcggtactta cgtcactctt ggcacgggga
     2461 atccgcgttc caatgcaccg ttcccggccg cggaggctgg atcggtcccg gtgtcttcta
     2521 tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac taaacgagct
     2581 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag
     2641 ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     2701 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg
     2761 tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa
     2821 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     2881 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     2941 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     3001 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     3061 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     3121 gtaattgatt actattaata actagtcaat aatcaatgcc aacatggcgg tcatattgga
     3181 catgagccaa tataaatgta catattatga tatagataca acgtatgcaa tggccaatag
     3241 ccaatattga tttatgctat ataaccaatg aataatatgg ctaatggcca atattgagtc
     3301 aacgccactg tgcatgaagc tcgtaattgt tatccgctca caattaaggg cgacacaaaa
     3361 tttattctaa atgataataa atactgataa catcttatag tttgtattat attttgtatt
     3421 atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta ttttcgagat
     3481 ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa atcataaata
     3541 atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta tcttatttaa
     3601 agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca agcaaagtga
     3661 caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca taaaaaaacc
     3721 cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc agaaccgccc
     3781 agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt tgtagaaacg
     3841 caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt tccctactct
     3901 cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta actcacgtta
     4741 agggattttg gtcatgagca aggggtgtta tgagccatat tcaggtataa atgggctcgc
     4801 gataatgttc agaattggtt aattggttgt aacactgacc cctatttgtt tatttttcta
     4861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4921 ttgaaaaagg aagaatatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4981 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5041 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5101 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5161 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5221 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5281 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5341 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5401 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5461 gcgtgacacc acgatgcctg tagcgatggc aacaacgttg cgcaaactat taactggcga
     5521 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5581 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatccggagc
     5641 cggtgagcgt ggttctcgcg gtatcatcgc agcgctgggg ccagatggta agccctcccg
     5701 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5761 cgctgagata ggtgcctcac tgattaagca ttggtaatga ccatgattac gccaagctct
     5821 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
     5881 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     5941 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
     6001 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     6061 catagaaggc ggcggtggaa tcgaaatctc gtagcacgtg tcaggcaccg ggcttgcggg
     6121 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
     6181 gccgctcgta gaaggggagg ttgcggggcg cggatgtctc caggaaggcg ggcaccccgg
     6241 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
     6301 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
     6361 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
     6421 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
     6481 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
     6541 ggaagagttc ttgcagctcg gtgacccgct cgat
//
