LOCUS       sequence-405788-        7435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            Fc(DAPA)-Avi-6xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7435)
  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 7435)
  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-IGSF21-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7435
                     /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             3011..3055
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3056..3073
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3213..3230)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3219..3330
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3448..3468)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3453..3782
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3633..3768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3695..3714
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3830..3877
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3896..4495
                     /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(3896..3915)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4392..4412
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4625..4746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4662..4681)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4716..4735
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4799..5590,5591..5659))
                     /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     5422..5441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5660..5751)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5793..5812
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5818..5837
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5923..6511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6412..6431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6665..6682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6780..6797
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6781..6797
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7253..7269
                     /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 cctatctgac cgtgaatatc gagcctctgc ctcctgtggt
     1021 ggctggcgac gctgtgaccc tgaagtgcaa cttcaagacc gacggccgga tgcgcgagat
     1081 cgtgtggtac agagttaccg acggcggcac catcaagcag aagatcttta ccttcgacgc
     1141 catgttcagc accaactaca gccacatgga aaactaccgg aagcgcgagg acctggtgta
     1201 ccagagcaca gtcagactgc ccgaagtgcg gatcagcgac aatggccctt acgagtgtca
     1261 cgtgggcatc tacgaccggg ccaccagaga aaaagtggtg ctggcctccg gcaacatctt
     1321 cctgaatgtg atggcccctc ctaccagcat cgaagtggtg gccgctgata cccctgctcc
     1381 tttcagcaga tatcaggccc agaacttcac cctcgtgtgc atcgtgtctg gcggcaagcc
     1441 tgctcctatg gtgtacttca agagagatgg cgagcccatc gacgccgtgc ctttgtctga
     1501 acctcctgcc gcttctagcg gccctctgca agacagcaga cccttcagaa gcctgctgca
     1561 cagagatctg gacgacacca agatgcagaa gtccctgagc ctgctggacg ccgagaatag
     1621 aggcggcaga ccctacaccg aaagacccag cagaggcctg acacctgatc ctaacatcct
     1681 gctgcagccc accaccgaga acatccctga gacagtggtg tcccgcgagt tccctagatg
     1741 ggtgcactct gccgagccta cctacttcct gcggcacagc agaaccccta gctctgatgg
     1801 cacagtggaa gtgcgggctc tgctgacctg gacactgaac cctcagatcg acaacgaggc
     1861 cctgttcagc tgcgaagtga agcaccctgc tctgagcatg cccatgcagg ccgaagtgac
     1921 actggtggct cctaagggcc ccaagatcgt gatgacaccc tccagagcca gagtgggcga
     1981 taccgtcaga atcctggtgc acggcttcca gaacgaggtg ttccccgagc ctatgttcac
     2041 ctggacaaga gtgggcagca gactgctgga tggcagcgcc gagttcgacg gcaaagaact
     2101 ggtgctggaa agagtgcccg ccgagctgaa tggcagcatg tacagatgta cagcccagaa
     2161 tcctctgggc agcaccgaca cacacacccg gctgatcgtg ttcgagaacc caaacatccc
     2221 cagaggcacc gaggacagca atggctctat cggacctaca ggcgccagac tgacacttgt
     2281 gctggccctg acagtgatcc tggaactgac aggcgcgccg ggcggcgaca agacccacac
     2341 ctgccccccc tgcccagctc cagaactgct gggagggcct tccgtgttcc tgttcccccc
     2401 caagcccaag gacaccctga tgatcagcag gacccccgag gtgacctgcg tggtggtggc
     2461 cgtgtcccac gaggacccag aggtgaagtt caactggtac gtggacggcg tggaggtgca
     2521 caacgccaag accaagccca gagaggagca gtacaacagc acctacaggg tggtgtccgt
     2581 gctgaccgtg ctgcaccagg actggctgaa cggcaaagaa tacaagtgca aagtctccaa
     2641 caaggccctg gctgccccaa tcgaaaagac aatcagcaag gccaagggcc agccacggga
     2701 gccccaggtg tacaccctgc cccccagccg ggaggagatg accaagaacc aggtgtccct
     2761 gacctgtctg gtgaagggct tctaccccag cgatatcgcc gtggagtggg agagcaacgg
     2821 ccagcccgag aacaactaca agaccacccc cccagtgctg gacagcgacg gcagcttctt
     2881 cctgtacagc aagctgaccg tggacaagtc caggtggcag cagggcaacg tgttcagctg
     2941 cagcgtgatg cacgaggccc tgcacaacca ctacacccag aagtccctga gcctgagccc
     3001 cggcggcagc ggtcttaatg atattttcga ggctcagaag attgagtggc atgaacacca
     3061 ccaccatcac cactaaggcc ggcctgtagt cactggcaca acgcgttacc cggtaagcca
     3121 atcgggtata cacggtcgtc atactgcaga cagggttctt ctactttgca agatagtctt
     3181 gagtagtaaa ataaatagat agagaaaatc agcctcgact gtgccttcta gttgccagcc
     3241 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3301 cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc ggtctattct
     3361 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3421 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3481 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3541 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3601 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3661 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3721 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3781 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     3841 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatgac
     3901 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3961 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     4021 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4081 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4141 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4201 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4261 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4321 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4381 atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4441 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacacgt
     4501 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4561 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4621 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4681 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4741 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcatt
     4801 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4861 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4921 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
     4981 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5041 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5101 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5161 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5221 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5281 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5341 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5401 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5461 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5521 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5581 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5641 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5701 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
     5761 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     5821 gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt
     5881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6061 aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact ctgtagcacc
     6121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6541 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6601 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6661 gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc caggcatcaa
     6721 actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc tttctgtgtt
     6781 gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa cgagtaatcg
     6841 ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg
     6901 gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata tatgagaatt
     6961 atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt tgctttttcg
     7021 attgatgaac acctataatt aaactattca tctattattt atgatttttt gtatatacaa
     7081 tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata aagggaaaat
     7141 cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata
     7201 agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt aattgtgagc
     7261 ggataacaat tacgagcttc atgcacagtg gcgttgactc aatattggcc attagccata
     7321 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     7381 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
