LOCUS       sequence-424381-        6649 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 6649)
  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 6649)
  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-FCGR2C-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2225..2269
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2270..2287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2427..2444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2433..2544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2662..2682)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2667..2996
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2847..2982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2909..2928
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3044..3091
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3110..3709
                     /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(3110..3129)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3606..3626
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3839..3960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3876..3895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3930..3949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4013..4804,4805..4873))
                     /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     4636..4655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4874..4965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5007..5026
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5032..5051
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5137..5725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5626..5645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5879..5896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5994..6011
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5995..6011
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6467..6483
                     /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)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg ccacaccagc tgctcctcca aaagctgtgc tgaagctgga
     1021 accccagtgg atcaacgtgc tgcaagagga tagcgtgacc ctgacctgta gaggcacaca
     1081 cagccctgag agcgacagca tccagtggtt ccacaacggc aatctgatcc ccacacacac
     1141 ccagcctagc taccggttca aggccaacaa caacgacagc ggcgagtaca cctgtcagac
     1201 aggccagaca tctctgagcg atcctgtgca cctgacagtg ctgtctgaat ggctggtgct
     1261 gcagacccct cacctggaat ttcaagaggg cgagacaatc gtgctgcggt gccactcttg
     1321 gaaggacaag cctctggtca aagtgacatt cttccagaac gggaagtcca agaagttcag
     1381 cagaagcgac cccaacttca gcatccctca ggccaatcac agccactccg gcgattacca
     1441 ctgtaccggc aatatcggct acaccctgta cagcagcaag cccgtgacaa tcacagtgca
     1501 ggcccctagc agcagcccta tgggaggcgc gccgggcggc gacaagaccc acacctgccc
     1561 cccctgccca gctccagaac tgctgggagg gccttccgtg ttcctgttcc cccccaagcc
     1621 caaggacacc ctgatgatca gcaggacccc cgaggtgacc tgcgtggtgg tggccgtgtc
     1681 ccacgaggac ccagaggtga agttcaactg gtacgtggac ggcgtggagg tgcacaacgc
     1741 caagaccaag cccagagagg agcagtacaa cagcacctac agggtggtgt ccgtgctgac
     1801 cgtgctgcac caggactggc tgaacggcaa agaatacaag tgcaaagtct ccaacaaggc
     1861 cctggctgcc ccaatcgaaa agacaatcag caaggccaag ggccagccac gggagcccca
     1921 ggtgtacacc ctgcccccca gccgggagga gatgaccaag aaccaggtgt ccctgacctg
     1981 tctggtgaag ggcttctacc ccagcgatat cgccgtggag tgggagagca acggccagcc
     2041 cgagaacaac tacaagacca cccccccagt gctggacagc gacggcagct tcttcctgta
     2101 cagcaagctg accgtggaca agtccaggtg gcagcagggc aacgtgttca gctgcagcgt
     2161 gatgcacgag gccctgcaca accactacac ccagaagtcc ctgagcctga gccccggcgg
     2221 cagcggtctt aatgatattt tcgaggctca gaagattgag tggcatgaac accaccacca
     2281 tcaccactaa ggccggcctg tagtcactgg cacaacgcgt tacccggtaa gccaatcggg
     2341 tatacacggt cgtcatactg cagacagggt tcttctactt tgcaagatag tcttgagtag
     2401 taaaataaat agatagagaa aatcagcctc gactgtgcct tctagttgcc agccatctgt
     2461 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2521 ctaataaaat gaggaaattg catcacaaca ctcaacccta tctcggtcta ttcttttgat
     2581 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2641 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2701 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2761 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2821 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2881 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2941 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3001 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     3061 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tgaccgagta
     3121 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     3181 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     3241 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     3301 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     3361 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     3421 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     3481 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     3541 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacatccgc
     3601 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     3661 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac acgtgctacg
     3721 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3781 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3841 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3901 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3961 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt cattaccaat
     4021 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4081 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagcgctg
     4141 cgatgatacc gcgagaacca cgctcaccgg ctccggattt atcagcaata aaccagccag
     4201 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4261 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4321 ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4381 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4441 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4501 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4561 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4621 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4681 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4741 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4801 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4861 gttgaatact catattcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4921 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gtcagtgtta
     4981 caaccaatta accaattctg aacattatcg cgagcccatt tatacctgaa tatggctcat
     5041 aacacccctt gctcatgacc aaaatccctt aacgtgagtt acgcgcgcgt cgttccactg
     5101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5281 tgttcttcta gtgtagccgt agttagccca ccacttcaag aactctgtag caccgcctac
     5341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5761 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5821 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5881 cgagtcagtg agcgaggaag cggaaggcga gagtagggaa ctgccaggca tcaaactaag
     5941 cagaaggccc ctgacggatg gcctttttgc gtttctacaa actctttctg tgttgtaaaa
     6001 cgacggccag tcttaagctc gggccccctg ggcggttctg ataacgagta atcgttaatc
     6061 cgcaaataac gtaaaaaccc gcttcggcgg gtttttttat ggggggagtt tagggaaaga
     6121 gcatttgtca gaatatttaa gggcgcctgt cactttgctt gatatatgag aattatttaa
     6181 ccttataaat gagaaaaaag caacgcactt taaataagat acgttgcttt ttcgattgat
     6241 gaacacctat aattaaacta ttcatctatt atttatgatt ttttgtatat acaatatttc
     6301 tagtttgtta aagagaatta agaaaataaa tctcgaaaat aataaaggga aaatcagttt
     6361 ttgatatcaa aattatacat gtcaacgata atacaaaata taatacaaac tataagatgt
     6421 tatcagtatt tattatcatt tagaataaat tttgtgtcgc ccttaattgt gagcggataa
     6481 caattacgag cttcatgcac agtggcgttg actcaatatt ggccattagc catattattc
     6541 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat
     6601 cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgtt
//
