LOCUS       sequence-368750-        6646 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 6646)
  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 6646)
  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-FCGR3B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     238..258
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    376..487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     476..493
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(633..650)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(651..695)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2052..2071)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2084..2102)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2085..2102)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2312..2336)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2333..2536)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2362..2382)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2537..2916
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3083..3099
                     /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)."
     primer_bind     complement(3555..3572)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3555..3571)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3670..3687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3841..4429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3921..3940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4515..4534)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4540..4559)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4601..4692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4693..4761,4762..5553)
                     /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     complement(4911..4930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5606..5727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5617..5636)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5671..5690
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5857..6456)
                     /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(5940..5960)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6437..6456
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6475..6522)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6570..6646,1..253))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6584..6646,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(6638..6646,1..11))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
       61 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
      121 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
      181 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
      241 ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt
      301 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      361 agggttgagt gttgtgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
      421 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
      481 ggcacagtcg aggctgattt tctctatcta tttattttac tactcaagac tatcttgcaa
      541 agtagaagaa ccctgtctgc agtatgacga ccgtgtatac ccgattggct taccgggtaa
      601 cgcgttgtgc cagtgactac aggccggcct tagtggtgat ggtggtggtg ttcatgccac
      661 tcaatcttct gagcctcgaa aatatcatta agaccgctgc cgccggggct caggctcagg
      721 gacttctggg tgtagtggtt gtgcagggcc tcgtgcatca cgctgcagct gaacacgttg
      781 ccctgctgcc acctggactt gtccacggtc agcttgctgt acaggaagaa gctgccgtcg
      841 ctgtccagca ctgggggggt ggtcttgtag ttgttctcgg gctggccgtt gctctcccac
      901 tccacggcga tatcgctggg gtagaagccc ttcaccagac aggtcaggga cacctggttc
      961 ttggtcatct cctcccggct ggggggcagg gtgtacacct ggggctcccg tggctggccc
     1021 ttggccttgc tgattgtctt ttcgattggg gcagccaggg ccttgttgga gactttgcac
     1081 ttgtattctt tgccgttcag ccagtcctgg tgcagcacgg tcagcacgga caccaccctg
     1141 taggtgctgt tgtactgctc ctctctgggc ttggtcttgg cgttgtgcac ctccacgccg
     1201 tccacgtacc agttgaactt cacctctggg tcctcgtggg acacggccac caccacgcag
     1261 gtcacctcgg gggtcctgct gatcatcagg gtgtccttgg gcttgggggg gaacaggaac
     1321 acggaaggcc ctcccagcag ttctggagct gggcaggggg ggcaggtgtg ggtcttgtcg
     1381 ccgcccggcg cgccagacac ggcgaggccc tgtgtgatgg tgatgttcac tgtctcgctg
     1441 gacacgttct tgctgcccac gaggcctcta cagaaatagg agccggagtc cttcagtgtg
     1501 gccttgggga tgtgaaagtc gctgttgtgg tggaagtact ttctgtcctt gccgttctgc
     1561 aggtaggtca ctttgtgcag ggctgtgttc ttccaagagt ggcatctcag gtggatgggg
     1621 tcctcttctt tgaacaccca tctaggggcc tgcagcagca gccatccaat atgcacttcc
     1681 agctgcacag ggtcgctcag ggtgctcaga ttggtctggc atctgtactc gccgctgtcg
     1741 ttcactgtgg cggcgtcgat aaagtagctg ctggcctggc tgctgatcag gctctcgttg
     1801 tggaaccact gggtgctgtt atcctcggga gagtaggcgc cctgacactt cagggtcacg
     1861 ctgtcctttt ccagcacgct gtaccactga ggttccagga acaccacggc tttaggcaga
     1921 tcctctgttc tcatggcggc cgcagcgaac accaggcaga agatgtagct cagggcaatg
     1981 atggtcttca tggtggcgct agcaggcgcg ttttcgatag aaccgaggtg cagttggacc
     2041 tgggagtgga cacctgtgga gagaaaggca aagtggatgt tattctatag tgtcacctaa
     2101 atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct aacgaagcca
     2161 agggggtggg cctatagact ctataggcgg tacttacgtc actcttggca cggggaatcc
     2221 gcgttccaat gcaccgttcc cggccgcgga ggctggatcg gtcccggtgt cttctatgga
     2281 ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctctgc
     2341 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     2401 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     2461 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     2521 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     2581 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     2641 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     2701 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     2761 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     2821 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     2881 ttgattacta ttaataacta gtcaataatc aatgccaaca tggcggtcat attggacatg
     2941 agccaatata aatgtacata ttatgatata gatacaacgt atgcaatggc caatagccaa
     3001 tattgattta tgctatataa ccaatgaata atatggctaa tggccaatat tgagtcaacg
     3061 ccactgtgca tgaagctcgt aattgttatc cgctcacaat taagggcgac acaaaattta
     3121 ttctaaatga taataaatac tgataacatc ttatagtttg tattatattt tgtattatcg
     3181 ttgacatgta taattttgat atcaaaaact gattttccct ttattatttt cgagatttat
     3241 tttcttaatt ctctttaaca aactagaaat attgtatata caaaaaatca taaataatag
     3301 atgaatagtt taattatagg tgttcatcaa tcgaaaaagc aacgtatctt atttaaagtg
     3361 cgttgctttt ttctcattta taaggttaaa taattctcat atatcaagca aagtgacagg
     3421 cgcccttaaa tattctgaca aatgctcttt ccctaaactc cccccataaa aaaacccgcc
     3481 gaagcgggtt tttacgttat ttgcggatta acgattactc gttatcagaa ccgcccaggg
     3541 ggcccgagct taagactggc cgtcgtttta caacacagaa agagtttgta gaaacgcaaa
     3601 aaggccatcc gtcaggggcc ttctgcttag tttgatgcct ggcagttccc tactctcgcc
     3661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtgggc
     4261 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4441 gatcttttct acggggtctg acgctcagtg gaacgacgcg cgcgtaactc acgttaaggg
     4501 attttggtca tgagcaaggg gtgttatgag ccatattcag gtataaatgg gctcgcgata
     4561 atgttcagaa ttggttaatt ggttgtaaca ctgaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4921 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5221 gacaccacga tgcctgtagc gatggcaaca acgttgcgca aactattaac tggcgaacta
     5281 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc cggagccggt
     5401 gagcgtggtt ctcgcggtat catcgcagcg ctggggccag atggtaagcc ctcccgtatc
     5461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5521 gagataggtg cctcactgat taagcattgg taatgaccat gattacgcca agctctagct
     5581 agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact
     5641 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     5701 accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc
     5761 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     5821 gaaggcggcg gtggaatcga aatctcgtag cacgtgtcag gcaccgggct tgcgggtcat
     5881 gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg
     5941 ctcgtagaag gggaggttgc ggggcgcgga tgtctccagg aaggcgggca ccccggcgcg
     6001 ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc
     6061 gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag
     6121 gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg
     6181 gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac
     6241 cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa
     6301 gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt
     6361 ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg
     6421 ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt tagttcctca ccttgtcgta
     6481 ttatactatg ccgatatact atgccgatga ttaattgtca acacgtgctg atcagatccg
     6541 aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     6601 cctcactact tctggaatag ctcagaggca gaggcggcct cggcct
//
