LOCUS       sequence-417860-        7270 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 7270)
  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 7270)
  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-FCGRT-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7270
                     /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             2846..2890
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2891..2908
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3048..3065)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3054..3165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3283..3303)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3288..3617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3468..3603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3530..3549
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3665..3712
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3731..4330
                     /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(3731..3750)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4227..4247
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4460..4581
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4497..4516)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4551..4570
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4634..5425,5426..5494))
                     /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     5257..5276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5495..5586)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5628..5647
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5653..5672
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5758..6346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6247..6266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6500..6517
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6615..6632
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6616..6632
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7088..7104
                     /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 ccattcagag aacccctaag atccaggtgt acagcagaca
     1021 ccccgccgag aacggcaaga gcaacttcct gaactgctac gtgtccggct ttcaccccag
     1081 cgacattgag gtggacctgc tgaagaacgg cgagcggatc gagaaggtgg aacacagcga
     1141 tctgagcttc agcaaggact ggtccttcta cctgctgtac tacaccgagt tcacccctac
     1201 cgagaaggac gagtacgcct gcagagtgaa ccacgtgaca ctgagccagc ctaagatcgt
     1261 gaagtgggac agagacatgg gcggaggcgg atctggtggc ggaggaagtg gcggcggagg
     1321 atctgctgaa tctcacctga gcctgctgta tcatctgacc gccgtgtcta gccccgctcc
     1381 tggaacacca gctttttggg tttccggctg gctgggccct cagcagtacc tgagctacaa
     1441 tagcctgaga ggcgaggccg aaccttgtgg cgcttgggtc tgggaaaatc aggtgtcctg
     1501 gtactgggag aaagagacaa ccgacctgcg gatcaaagag aagctgttcc tggaagcctt
     1561 caaggccctc ggcggaaagg gcccatatac actgcaaggc ctgctgggct gtgaactggg
     1621 ccctgataat accagcgtgc ccaccgccaa gttcgccctg aatggggaag agttcatgaa
     1681 tttcgacctg aagcaaggca cctggggcgg agattggcct gaagctctgg ccattagcca
     1741 gagatggcag cagcaggata aggccgccaa caaagagctg accttcctgc tgttcagctg
     1801 ccctcacaga ctgagagagc acctggaaag aggcagaggc aacctggaat ggaaagaacc
     1861 tcctagcatg cggctgaagg ccagacctag cagccctggc tttagcgtgc tgacctgtag
     1921 cgccttcagc ttctaccctc ctgaactcca gctgcggttc ctgagaaatg gactggccgc
     1981 tggaacaggc cagggcgatt ttggccctaa tagcgacggc agctttcacg cctctagcag
     2041 cctgacagtg aagtccggcg acgagcacca ctactgctgc attgtgcagc atgccggact
     2101 ggcccagcct ctgagagtgg aacttgagag ccctgccaag tcctctggcg cgccgggcgg
     2161 cgacaagacc cacacctgcc ccccctgccc agctccagaa ctgctgggag ggccttccgt
     2221 gttcctgttc ccccccaagc ccaaggacac cctgatgatc agcaggaccc ccgaggtgac
     2281 ctgcgtggtg gtggccgtgt cccacgagga cccagaggtg aagttcaact ggtacgtgga
     2341 cggcgtggag gtgcacaacg ccaagaccaa gcccagagag gagcagtaca acagcaccta
     2401 cagggtggtg tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aagaatacaa
     2461 gtgcaaagtc tccaacaagg ccctggctgc cccaatcgaa aagacaatca gcaaggccaa
     2521 gggccagcca cgggagcccc aggtgtacac cctgcccccc agccgggagg agatgaccaa
     2581 gaaccaggtg tccctgacct gtctggtgaa gggcttctac cccagcgata tcgccgtgga
     2641 gtgggagagc aacggccagc ccgagaacaa ctacaagacc acccccccag tgctggacag
     2701 cgacggcagc ttcttcctgt acagcaagct gaccgtggac aagtccaggt ggcagcaggg
     2761 caacgtgttc agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc
     2821 cctgagcctg agccccggcg gcagcggtct taatgatatt ttcgaggctc agaagattga
     2881 gtggcatgaa caccaccacc atcaccacta aggccggcct gtagtcactg gcacaacgcg
     2941 ttacccggta agccaatcgg gtatacacgg tcgtcatact gcagacaggg ttcttctact
     3001 ttgcaagata gtcttgagta gtaaaataaa tagatagaga aaatcagcct cgactgtgcc
     3061 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3121 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct
     3181 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3241 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3301 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3361 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3421 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3481 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3541 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3601 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     3661 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     3721 gaactaaacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     3781 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     3841 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     3901 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     3961 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     4021 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     4081 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     4141 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     4201 cgccttcctg gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     4261 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     4321 cggtgcctga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4381 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4441 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4501 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4561 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4621 gtaatcatgg tcattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4681 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4741 taccatctgg ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt
     4801 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4861 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4921 atagtttgcg caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4981 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5041 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5101 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5161 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5221 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5281 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5341 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5401 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5461 gaataagggc gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa
     5521 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5581 aacaaatagg ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat
     5641 ttatacctga atatggctca taacacccct tgctcatgac caaaatccct taacgtgagt
     5701 tacgcgcgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5761 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5821 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5881 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa
     5941 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6001 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6061 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6121 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6181 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6241 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6301 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6361 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6421 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6481 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga
     6541 actgccaggc atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca
     6601 aactctttct gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct
     6661 gataacgagt aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta
     6721 tggggggagt ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct
     6781 tgatatatga gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga
     6841 tacgttgctt tttcgattga tgaacaccta taattaaact attcatctat tatttatgat
     6901 tttttgtata tacaatattt ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa
     6961 taataaaggg aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat
     7021 ataatacaaa ctataagatg ttatcagtat ttattatcat ttagaataaa ttttgtgtcg
     7081 cccttaattg tgagcggata acaattacga gcttcatgca cagtggcgtt gactcaatat
     7141 tggccattag ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca
     7201 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
     7261 ccgccatgtt
//
