LOCUS       sequence-417400-        7075 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 7075)
  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 7075)
  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-PVR-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7075
                     /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             2651..2695
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2696..2713
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2853..2870)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2859..2970
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3088..3108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3093..3422
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3273..3408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3335..3354
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3470..3517
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3536..4135
                     /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(3536..3555)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4032..4052
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4265..4386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4302..4321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4356..4375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4439..5230,5231..5299))
                     /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     5062..5081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5300..5391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5433..5452
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5458..5477
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5563..6151
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6052..6071
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6305..6322
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6420..6437
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6421..6437
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6893..6909
                     /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 cctggcctcc acctggaaca ggggatgttg ttgttcaggc
     1021 ccctacacag gtgcccggct ttctgggaga ttctgtgacc ctgccttgct acctgcaagt
     1081 gcccaacatg gaagtgaccc acgtgtccca gctgacatgg gccagacatg gcgaatctgg
     1141 cagcatggcc gtgtttcacc agacacaggg ccctagctac agcgagagca agagactgga
     1201 attcgtggcc gccagactgg gcgctgaact gagaaatgcc agcctgagaa tgttcggcct
     1261 gagagtggaa gatgagggca actacacctg tctgttcgtg acctttccac agggcagcag
     1321 aagcgtggac atctggctga gagtgctggc caaacctcag aataccgccg aggtgcagaa
     1381 agtgcagctg acaggcgaac ctgtgcctat ggccagatgt gtgtctactg gtggcagacc
     1441 tcctgctcag atcacctggc attctgatct cggcggcatg cccaatacca gccaggttcc
     1501 aggatttctg agcggcaccg tgacagtgac cagcctgtgg attctggtgc ctagctctca
     1561 ggtggacggc aagaatgtga cctgcaaggt ggaacacgag agcttcgaga agcctcagct
     1621 gctgaccgtg aacctgaccg tgtactatcc tcctgaggtg tccatcagcg gctacgacaa
     1681 caactggtat ctgggccaga acgaggccac actgacctgc gacgccagaa gcaatcctga
     1741 gcctaccggc tacaactggt ccaccacaat gggccctctg cctccatttg ctgttgctca
     1801 aggcgcccag ctgctcatca gacccgtgga caagcccatc aacacaaccc tgatctgcaa
     1861 cgtgacaaac gccctgggag ccagacaggc tgagctgaca gtgcaagtga aagagggacc
     1921 tcctagcgag cacagcggca tcagcagaaa tggcgcgccg ggcggcgaca agacccacac
     1981 ctgccccccc tgcccagctc cagaactgct gggagggcct tccgtgttcc tgttcccccc
     2041 caagcccaag gacaccctga tgatcagcag gacccccgag gtgacctgcg tggtggtggc
     2101 cgtgtcccac gaggacccag aggtgaagtt caactggtac gtggacggcg tggaggtgca
     2161 caacgccaag accaagccca gagaggagca gtacaacagc acctacaggg tggtgtccgt
     2221 gctgaccgtg ctgcaccagg actggctgaa cggcaaagaa tacaagtgca aagtctccaa
     2281 caaggccctg gctgccccaa tcgaaaagac aatcagcaag gccaagggcc agccacggga
     2341 gccccaggtg tacaccctgc cccccagccg ggaggagatg accaagaacc aggtgtccct
     2401 gacctgtctg gtgaagggct tctaccccag cgatatcgcc gtggagtggg agagcaacgg
     2461 ccagcccgag aacaactaca agaccacccc cccagtgctg gacagcgacg gcagcttctt
     2521 cctgtacagc aagctgaccg tggacaagtc caggtggcag cagggcaacg tgttcagctg
     2581 cagcgtgatg cacgaggccc tgcacaacca ctacacccag aagtccctga gcctgagccc
     2641 cggcggcagc ggtcttaatg atattttcga ggctcagaag attgagtggc atgaacacca
     2701 ccaccatcac cactaaggcc ggcctgtagt cactggcaca acgcgttacc cggtaagcca
     2761 atcgggtata cacggtcgtc atactgcaga cagggttctt ctactttgca agatagtctt
     2821 gagtagtaaa ataaatagat agagaaaatc agcctcgact gtgccttcta gttgccagcc
     2881 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2941 cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc ggtctattct
     3001 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3061 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3241 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3301 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3361 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3421 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     3481 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatgac
     3541 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3601 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3661 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3721 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3781 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3841 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     3901 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     3961 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4021 atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4081 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacacgt
     4141 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4201 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4261 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4321 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4381 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcatt
     4441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4561 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
     4621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4741 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5281 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
     5401 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     5461 gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt
     5521 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5581 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5641 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5701 aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact ctgtagcacc
     5761 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5821 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5881 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5941 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6001 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6061 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6121 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6181 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6241 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6301 gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc caggcatcaa
     6361 actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc tttctgtgtt
     6421 gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa cgagtaatcg
     6481 ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg ggagtttagg
     6541 gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata tatgagaatt
     6601 atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt tgctttttcg
     6661 attgatgaac acctataatt aaactattca tctattattt atgatttttt gtatatacaa
     6721 tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata aagggaaaat
     6781 cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat acaaactata
     6841 agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt aattgtgagc
     6901 ggataacaat tacgagcttc atgcacagtg gcgttgactc aatattggcc attagccata
     6961 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     7021 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgtt
//
