LOCUS       sequence-401035-        8905 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 8905)
  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 8905)
  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-IGDCC4-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8905
                     /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             4481..4525
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4526..4543
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4683..4700)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4689..4800
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4918..4938)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4923..5252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5103..5238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5165..5184
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5300..5347
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5366..5965
                     /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(5366..5385)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5862..5882
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6095..6216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6132..6151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6186..6205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6269..7060,7061..7129))
                     /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     6892..6911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7130..7221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7263..7282
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7288..7307
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7393..7981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7882..7901
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8135..8152
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8250..8267
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8251..8267
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8723..8739
                     /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 ccgaattgct gctgcctcaa gagacaaccg tggaactgag
     1021 ctgtggcgtg ggacccctgc aagtgattct gggacctgaa caggccgccg tgctgaattg
     1081 ctctcttgga gctgctgctg caggccctcc taccagagtg acctggtcta aggatggcga
     1141 caccctgctg gaacacgacc atctgcatct gctgcccaat ggcagcctgt ggctgtctca
     1201 acccctggct cctaatggct ccgatgagtc tgtgcctgaa gccgtgggag tgatcgaggg
     1261 caactacagc tgtctggccc atggacctct gggagtgctg gcttctcaga cagccgtggt
     1321 taagctggcc acactggccg actttagtct gcaccctgag agccagaccg tggaagagaa
     1381 tggcaccgcc agattcgagt gccacatcga gggactgccc gctcctatca tcacctggga
     1441 gaaagaccaa gtgaccctgc ctgaggaacc ccggctgatc gttctgccta atggcgtgct
     1501 gcagatcctg gacgtgcaag agtctgacgc tggcccctat agatgcgtgg ccaccaatag
     1561 cgccaggcag cacttttctc aagaggccct gctgagcgtg gcccacagag gatctctggc
     1621 ctctacaaga ggccaggacg tggtcattgt ggctgcccct gagaatacca ccgtggtgtc
     1681 tggccagtcc gtggtcatgg aatgtgtggc ctctgccgat cctacacctt tcgtgtcctg
     1741 ggtccgacag gacggcaagc ctatcagcac cgatgtgatc gtgctgggcc gcaccaatct
     1801 gctgatcgcc aatgctcagc cttggcactc tggcgtgtac gtgtgcagag ccaacaagcc
     1861 cagaaccaga gactttgcca cagccgccgc tgagctgaga gttcttgctg ccccagccat
     1921 tacacaggcc cctgaagctc tgagcagaac cagggcttcc acagccagat ttgtgtgtag
     1981 agccagcggc gagcctcggc ctgctcttag atggcttcat aatggcgccc ctctgaggcc
     2041 caacggcaga gtgaaagttc aaggcggagg cggcagcctg gtcatcacac agattggact
     2101 gcaggacgcc ggctactacc agtgcgtggc cgaaaattct gccggcatgg cttgtgccgc
     2161 tgcttctctg gcagtggtcg ttagagaagg cctgcctagc gctcccacaa gagtgacagc
     2221 tacaccactg agcagctctg ccgtgctggt ggcttgggaa agacctgaga tgcacagcga
     2281 gcagatcatc ggcttcagcc tgcactacca gaaagccaga ggcatggaca acgtggaata
     2341 ccagttcgcc gtgaacaacg acaccaccga gctgcaagtg cgcgacctgg aacctaacac
     2401 cgactacgag ttctacgtgg tggcctacag tcagctgggc gccagcagaa catctacccc
     2461 tgctctggtg cacaccctgg acgatgttcc ttctgctgct ccacagctga gcctgagcag
     2521 ccccaatcct agcgatatca gagtggcctg gctgcctctg cctccttctc tgtctaatgg
     2581 ccaggtggtc aagtacaaga tcgagtacgg cctgggcaaa gaggaccaga tcttctccac
     2641 agaagtgcgg ggcaacgaga cacagctgat gctcaatagc ctgcagccta acaaggtgta
     2701 cagagtgcgg atcagcgccg gaacagccgc tggatttgga gcaccttctc agtggatgca
     2761 ccacagaacc cctagcatgc acaaccagag ccacgtgcca tttgctcccg ccgaactgaa
     2821 ggtgcaggcc aagatggaaa gcctggtggt gtcttggcag cctcctccac atcctacaca
     2881 gatctccggc tacaagctgt attggagaga agtgggcgcc gaggaagagg ccaacggcga
     2941 tagacttcct ggcggaagag gcgatcaggc ctgggatgtt ggacctgtgc ggctgaagaa
     3001 aaaagtgaag cagtacgagc tgacccagct ggtgcctggc agactgtacg aagtgaagct
     3061 ggtggccttc aacaagcacg aggatggcta tgccgccgtg tggaagggca agacagaaaa
     3121 ggctcccgct cctgacatgc ccatccagag aggacctcca ctgcctccag ctcatgtgca
     3181 cgccgagagc aatagcagca ccagcatctg gctgcggtgg aagaagcccg atttcaccac
     3241 cgtgaagatc gtgaactaca ccgtgcggtt cagcccctgg ggactgagaa atgcctctct
     3301 ggtcacctac tacaccagct ccggcgagga catcctgatc ggaggactga agcccttcac
     3361 caaatacgag tttgccgtgc agtcccacgg cgtggacatg gatggacctt ttggcagcgt
     3421 ggtggaaaga agcaccctgc cagacagacc cagcacacct ccaagcgacc tgagactgtc
     3481 tcccctgaca ccatctacag tgcggctgca ctggtgccct ccaacagagc caaatggcga
     3541 gatcgtggaa tatctgatcc tgtacagcag caaccacaca cagcccgagc accagtggac
     3601 cctgcttacc acacagggca acatcttcag cgccgaggtt cacggcctgg aaagcgatac
     3661 ccggtacttc ttcaagatgg gagccagaac cgaagtcggc cctggacctt tcagcagact
     3721 ccaggatgtg atcaccctgc aagagaagct gagcgacagc ctggacatgc actccgtgac
     3781 aggcgcgccg ggcggcgaca agacccacac ctgccccccc tgcccagctc cagaactgct
     3841 gggagggcct tccgtgttcc tgttcccccc caagcccaag gacaccctga tgatcagcag
     3901 gacccccgag gtgacctgcg tggtggtggc cgtgtcccac gaggacccag aggtgaagtt
     3961 caactggtac gtggacggcg tggaggtgca caacgccaag accaagccca gagaggagca
     4021 gtacaacagc acctacaggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa
     4081 cggcaaagaa tacaagtgca aagtctccaa caaggccctg gctgccccaa tcgaaaagac
     4141 aatcagcaag gccaagggcc agccacggga gccccaggtg tacaccctgc cccccagccg
     4201 ggaggagatg accaagaacc aggtgtccct gacctgtctg gtgaagggct tctaccccag
     4261 cgatatcgcc gtggagtggg agagcaacgg ccagcccgag aacaactaca agaccacccc
     4321 cccagtgctg gacagcgacg gcagcttctt cctgtacagc aagctgaccg tggacaagtc
     4381 caggtggcag cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca
     4441 ctacacccag aagtccctga gcctgagccc cggcggcagc ggtcttaatg atattttcga
     4501 ggctcagaag attgagtggc atgaacacca ccaccatcac cactaaggcc ggcctgtagt
     4561 cactggcaca acgcgttacc cggtaagcca atcgggtata cacggtcgtc atactgcaga
     4621 cagggttctt ctactttgca agatagtctt gagtagtaaa ataaatagat agagaaaatc
     4681 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4741 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4801 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4861 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4921 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4981 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5041 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5101 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5161 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5221 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5281 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     5341 atacgacaag gtgaggaact aaaccatgac cgagtacaag cccacggtgc gcctcgccac
     5401 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5461 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5521 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5581 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5641 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5701 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5761 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5821 gcgcgccggg gtgcccgcct tcctggagac atccgcgccc cgcaacctcc ccttctacga
     5881 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5941 catgacccgc aagcccggtg cctgacacgt gctacgagat ttcgattcca ccgccgcctt
     6001 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6061 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6121 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6181 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6241 tagctagagc ttggcgtaat catggtcatt accaatgctt aatcagtgag gcacctatct
     6301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6361 cgatacggga gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct
     6421 caccggctcc ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt
     6601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt
     7141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7201 gtatttagaa aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca
     7261 ttatcgcgag cccatttata cctgaatatg gctcataaca ccccttgctc atgaccaaaa
     7321 tcccttaacg tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     7381 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7441 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7501 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7561 agcccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7621 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7681 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7741 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7801 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7861 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7921 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7981 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8041 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     8101 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     8161 aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga cggatggcct
     8221 ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc
     8281 cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa aaacccgctt
     8341 cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat atttaagggc
     8401 gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga aaaaagcaac
     8461 gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt aaactattca
     8521 tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga gaattaagaa
     8581 aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt atacatgtca
     8641 acgataatac aaaatataat acaaactata agatgttatc agtatttatt atcatttaga
     8701 ataaattttg tgtcgccctt aattgtgagc ggataacaat tacgagcttc atgcacagtg
     8761 gcgttgactc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     8821 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     8881 tcatgtccaa tatgaccgcc atgtt
//
