LOCUS       sequence-401531-        6682 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 6682)
  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 6682)
  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-FCGR3A-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6682
                     /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             2258..2302
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2303..2320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2460..2477)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2466..2577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2695..2715)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2700..3029
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2880..3015
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2942..2961
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3077..3124
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3143..3742
                     /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(3143..3162)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3639..3659
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3872..3993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3909..3928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3963..3982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4046..4837,4838..4906))
                     /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     4669..4688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4907..4998)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5040..5059
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5065..5084
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5170..5758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5659..5678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5912..5929
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6027..6044
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6028..6044
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6500..6516
                     /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 ccggaatgag aacagaagat ctgcctaagg ccgtggtgtt
     1021 cctggaacct cagtggtaca gagtgctgga aaaggacagc gtgaccctga agtgtcaggg
     1081 cgcctactct cccgaggata acagcaccca gtggttccac aacgagagcc tgatcagcag
     1141 ccaggccagc agctacttta tcgatgccgc cacagtggac gacagcggcg agtacagatg
     1201 ccagaccaat ctgagcaccc tgagcgaccc tgtgcagctg gaagtgcata ttggatggct
     1261 gctgctgcag gcccctagat gggtgttcaa agaagaggac cccatccacc tgagatgcca
     1321 ctcttggaag aacacagccc tgcacaaagt gacctacctg cagaacggca agggcagaaa
     1381 gtacttccac cacaacagcg acttctacat ccccaaggcc acactgaagg actccggctc
     1441 ctatttctgc agaggcctgt tcggcagcaa gaacgtgtcc agcgagacag tgaacatcac
     1501 catcacacag ggcctcgccg tgtccaccat cagcagcttt tttccacctg gctaccaagg
     1561 cgcgccgggc ggcgacaaga cccacacctg ccccccctgc ccagctccag aactgctggg
     1621 agggccttcc gtgttcctgt tcccccccaa gcccaaggac accctgatga tcagcaggac
     1681 ccccgaggtg acctgcgtgg tggtggccgt gtcccacgag gacccagagg tgaagttcaa
     1741 ctggtacgtg gacggcgtgg aggtgcacaa cgccaagacc aagcccagag aggagcagta
     1801 caacagcacc tacagggtgg tgtccgtgct gaccgtgctg caccaggact ggctgaacgg
     1861 caaagaatac aagtgcaaag tctccaacaa ggccctggct gccccaatcg aaaagacaat
     1921 cagcaaggcc aagggccagc cacgggagcc ccaggtgtac accctgcccc ccagccggga
     1981 ggagatgacc aagaaccagg tgtccctgac ctgtctggtg aagggcttct accccagcga
     2041 tatcgccgtg gagtgggaga gcaacggcca gcccgagaac aactacaaga ccaccccccc
     2101 agtgctggac agcgacggca gcttcttcct gtacagcaag ctgaccgtgg acaagtccag
     2161 gtggcagcag ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta
     2221 cacccagaag tccctgagcc tgagccccgg cggcagcggt cttaatgata ttttcgaggc
     2281 tcagaagatt gagtggcatg aacaccacca ccatcaccac taaggccggc ctgtagtcac
     2341 tggcacaacg cgttacccgg taagccaatc gggtatacac ggtcgtcata ctgcagacag
     2401 ggttcttcta ctttgcaaga tagtcttgag tagtaaaata aatagataga gaaaatcagc
     2461 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2521 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcaca
     2581 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2641 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2701 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2761 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2821 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2881 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2941 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3001 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3061 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     3121 cgacaaggtg aggaactaaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3181 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3241 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3301 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3361 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3421 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3481 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3541 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3601 cgccggggtg cccgccttcc tggagacatc cgcgccccgc aacctcccct tctacgagcg
     3661 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     3721 gacccgcaag cccggtgcct gacacgtgct acgagatttc gattccaccg ccgccttcta
     3781 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3841 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3901 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3961 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4021 ctagagcttg gcgtaatcat ggtcattacc aatgcttaat cagtgaggca cctatctcag
     4081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4141 tacgggaggg cttaccatct ggccccagcg ctgcgatgat accgcgagaa ccacgctcac
     4201 cggctccgga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4321 gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac
     4381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatattc ttcctttttc
     4921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4981 tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa ttaaccaatt ctgaacatta
     5041 tcgcgagccc atttatacct gaatatggct cataacaccc cttgctcatg accaaaatcc
     5101 cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5161 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5221 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5281 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagc
     5341 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5401 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5461 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5521 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5581 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5641 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5701 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5761 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5821 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5881 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg
     5941 cgagagtagg gaactgccag gcatcaaact aagcagaagg cccctgacgg atggcctttt
     6001 tgcgtttcta caaactcttt ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc
     6061 ctgggcggtt ctgataacga gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg
     6121 cgggtttttt tatgggggga gtttagggaa agagcatttg tcagaatatt taagggcgcc
     6181 tgtcactttg cttgatatat gagaattatt taaccttata aatgagaaaa aagcaacgca
     6241 ctttaaataa gatacgttgc tttttcgatt gatgaacacc tataattaaa ctattcatct
     6301 attatttatg attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat
     6361 aaatctcgaa aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg
     6421 ataatacaaa atataataca aactataaga tgttatcagt atttattatc atttagaata
     6481 aattttgtgt cgcccttaat tgtgagcgga taacaattac gagcttcatg cacagtggcg
     6541 ttgactcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
     6601 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
     6661 tgtccaatat gaccgccatg tt
//
