LOCUS       sequence-471685-        7300 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 7300)
  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 7300)
  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-HLA-C-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             659..703
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             704..721
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(861..878)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    867..978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1096..1116)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1101..1430
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1281..1416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1343..1362
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1478..1525
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1544..2143
                     /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(1544..1563)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2040..2060
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2273..2394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2310..2329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2364..2383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(2447..3238,3239..3307))
                     /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     3070..3089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3308..3399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3441..3460
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3466..3485
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3571..4159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4060..4079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4313..4330
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4428..4445
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4429..4445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4901..4917
                     /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)."
     enhancer        5084..5463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5464..5667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5618..5638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5664..5688
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        5898..5916
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5898..5915
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     5929..5948
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 cccagctcca gaactgctgg gagggccttc cgtgttcctg ttccccccca agcccaagga
       61 caccctgatg atcagcagga cccccgaggt gacctgcgtg gtggtggccg tgtcccacga
      121 ggacccagag gtgaagttca actggtacgt ggacggcgtg gaggtgcaca acgccaagac
      181 caagcccaga gaggagcagt acaacagcac ctacagggtg gtgtccgtgc tgaccgtgct
      241 gcaccaggac tggctgaacg gcaaagaata caagtgcaaa gtctccaaca aggccctggc
      301 tgccccaatc gaaaagacaa tcagcaaggc caagggccag ccacgggagc cccaggtgta
      361 caccctgccc cccagccggg aggagatgac caagaaccag gtgtccctga cctgtctggt
      421 gaagggcttc taccccagcg atatcgccgt ggagtgggag agcaacggcc agcccgagaa
      481 caactacaag accacccccc cagtgctgga cagcgacggc agcttcttcc tgtacagcaa
      541 gctgaccgtg gacaagtcca ggtggcagca gggcaacgtg ttcagctgca gcgtgatgca
      601 cgaggccctg cacaaccact acacccagaa gtccctgagc ctgagccccg gcggcagcgg
      661 tcttaatgat attttcgagg ctcagaagat tgagtggcat gaacaccacc accatcacca
      721 ctaaggccgg cctgtagtca ctggcacaac gcgttacccg gtaagccaat cgggtataca
      781 cggtcgtcat actgcagaca gggttcttct actttgcaag atagtcttga gtagtaaaat
      841 aaatagatag agaaaatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
      901 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
      961 aaatgaggaa attgcatcac aacactcaac cctatctcgg tctattcttt tgatttataa
     1021 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1081 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1201 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1261 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1321 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1381 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     1441 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     1501 gtatatcggc atagtataat acgacaaggt gaggaactaa accatgaccg agtacaagcc
     1561 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     1621 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     1681 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     1741 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     1801 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     1861 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     1921 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     1981 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacat ccgcgccccg
     2041 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     2101 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacacgtgc tacgagattt
     2161 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     2221 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     2281 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2341 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2401 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcattac caatgcttaa
     2461 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2521 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga
     2581 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa
     2641 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2701 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     2761 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2821 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2881 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2941 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3001 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3061 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3121 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3181 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3241 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3301 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3361 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca
     3421 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc
     3481 ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc
     3541 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3601 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3661 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3721 tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc ctacatacct
     3781 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3841 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3901 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3961 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4021 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4081 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4141 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4201 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4261 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4321 agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac taagcagaag
     4381 gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg
     4441 ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt aatccgcaaa
     4501 taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga aagagcattt
     4561 gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat ttaaccttat
     4621 aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat tgatgaacac
     4681 ctataattaa actattcatc tattatttat gattttttgt atatacaata tttctagttt
     4741 gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca gtttttgata
     4801 tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag atgttatcag
     4861 tatttattat catttagaat aaattttgtg tcgcccttaa ttgtgagcgg ataacaatta
     4921 cgagcttcat gcacagtggc gttgactcaa tattggccat tagccatatt attcattggt
     4981 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     5041 atgtacattt atattggctc atgtccaata tgaccgccat gttggcattg attattgact
     5101 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5161 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5221 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5281 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5341 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5401 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5461 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5521 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5581 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5641 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     5701 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggccgg
     5761 gaacggtgca ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga
     5821 gtctataggc ccaccccctt ggcttcgtta gaacgcggct acaattaata cataacctta
     5881 tgtatcatac acatacgatt taggtgacac tatagaataa catccacttt gcctttctct
     5941 ccacaggtgt ccactcccag gtccaactgc acctcggttc tatcgaaaac gcgcctgcta
     6001 gcgccaccat gaagaccatc attgccctga gctacatctt ctgcctggtg ttcgctgcgg
     6061 ccgccattca gagaacccct aagatccagg tgtacagcag acaccccgcc gagaacggca
     6121 agagcaactt cctgaactgc tacgtgtccg gctttcaccc cagcgacatt gaggtggacc
     6181 tgctgaagaa cggcgagcgg atcgagaagg tggaacacag cgatctgagc ttcagcaagg
     6241 actggtcctt ctacctgctg tactacaccg agttcacccc taccgagaag gacgagtacg
     6301 cctgcagagt gaaccacgtg acactgagcc agcctaagat cgtgaagtgg gacagagaca
     6361 tgggcggagg cggatctggt ggcggaggaa gtggcggcgg aggatcttgt agccacagca
     6421 tgagatactt cgacaccgcc gtgtccagac ctggaagagg cgagcctaga ttcatcagcg
     6481 tgggctacgt ggacgacacc cagttcgtca gattcgacag cgacgccgct tctcccagag
     6541 gggaacctag agcaccttgg gttgagcaag agggccccga gtactgggat agagagacac
     6601 agaagtacaa gcggcaggcc caggccgata gagtgtccct gagaaacctg cggggctact
     6661 acaaccagag cgaggatggc tctcacaccc tgcagagaat gagcggctgt gatctgggcc
     6721 ctgatggcag actgctgaga ggctatgatc agagcgccta cgacggcaag gactatatcg
     6781 ccctgaacga ggacctgcgg tcttggacag ctgccgatac agctgcccag attacccagc
     6841 ggaaactgga agccgccaga gccgctgaac agctgagagc ttatctggaa ggcacctgtg
     6901 tggaatggct gcggagatac ctggaaaacg gcaaagagac actgcagcgg gccgagcctc
     6961 ctaagacaca tgtgacacac catcctctga gcgaccacga ggccacactg agatgttggg
     7021 ccctgggctt ttaccctgcc gagatcacac tgacctggca gagagatggc gaggatcaga
     7081 cccaggatac cgagctggtg gaaacaagac ctgccggcga cggcaccttc cagaaatggg
     7141 ctgctgtggt ggtgcctagc ggccaagagc agagatacac ctgtcacatg cagcacgagg
     7201 gcctgcaaga gcctctgaca ctgtcttggg agcctagcag ccagccaaca atccccattg
     7261 gcgcgccggg cggcgacaag acccacacct gccccccctg
//
