LOCUS       sequence-368744-        6631 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 6631)
  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 6631)
  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-FCGR2B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..216
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    334..445
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     434..451
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(591..608)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(609..653)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1995..2014)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2027..2045)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2028..2045)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2255..2279)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2276..2479)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2305..2325)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2480..2859
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3026..3042
                     /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(3498..3515)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3498..3514)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3613..3630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3784..4372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3864..3883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4458..4477)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4483..4502)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4544..4635
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4636..4704,4705..5496)
                     /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(4854..4873)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5549..5670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5560..5579)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5614..5633
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5800..6399)
                     /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(5883..5903)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6380..6399
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6418..6465)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6513..6631,1..211))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6527..6631,1..31)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6581..6600)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
       61 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      121 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      181 ctggggactt tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt
      241 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      301 aaaagaatag accgagatag ggttgagtgt tgtgatgcaa tttcctcatt ttattaggaa
      361 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
      421 gatggctggc aactagaagg cacagtcgag gctgattttc tctatctatt tattttacta
      481 ctcaagacta tcttgcaaag tagaagaacc ctgtctgcag tatgacgacc gtgtataccc
      541 gattggctta ccgggtaacg cgttgtgcca gtgactacag gccggcctta gtggtgatgg
      601 tggtggtgtt catgccactc aatcttctga gcctcgaaaa tatcattaag accgctgccg
      661 ccggggctca ggctcaggga cttctgggtg tagtggttgt gcagggcctc gtgcatcacg
      721 ctgcagctga acacgttgcc ctgctgccac ctggacttgt ccacggtcag cttgctgtac
      781 aggaagaagc tgccgtcgct gtccagcact gggggggtgg tcttgtagtt gttctcgggc
      841 tggccgttgc tctcccactc cacggcgata tcgctggggt agaagccctt caccagacag
      901 gtcagggaca cctggttctt ggtcatctcc tcccggctgg ggggcagggt gtacacctgg
      961 ggctcccgtg gctggccctt ggccttgctg attgtctttt cgattggggc agccagggcc
     1021 ttgttggaga ctttgcactt gtattctttg ccgttcagcc agtcctggtg cagcacggtc
     1081 agcacggaca ccaccctgta ggtgctgttg tactgctcct ctctgggctt ggtcttggcg
     1141 ttgtgcacct ccacgccgtc cacgtaccag ttgaacttca cctctgggtc ctcgtgggac
     1201 acggccacca ccacgcaggt cacctcgggg gtcctgctga tcatcagggt gtccttgggc
     1261 ttggggggga acaggaacac ggaaggccct cccagcagtt ctggagctgg gcaggggggg
     1321 caggtgtggg tcttgtcgcc gcccggcgcg cctggagcct gcactgtgat tgtcacgggc
     1381 ttgctgctgt acagggtgta gccgatattg ccggtacagt ggtaatcgcc ggagtggctg
     1441 tgattggcct gagggatgct gaagttgggg tcgcttctgc tgaacttctt ggacttcccg
     1501 ttctggaaga atgtcacttt gaccagaggc ttgtccttcc aagagtggca ccgcagcacg
     1561 attgtctcgc cctcttgaaa ttccaggtga ggggtctgca gcaccagcca ttcagacagc
     1621 actgtcaggt gcacaggatc gctcagagat gtctggcctg tctgacaggt gtactcgccg
     1681 ctgtcgttgt tgttggcctt gaaccggtag ctaggctggg tgtgtgtggg gatcagattg
     1741 ccgttgtgga accactggat gctgtcgctc tcagggctgt gtgtgcctct acaggtcagg
     1801 gtcacgctat cctcttgcag cacgttgatc cactggggtt ccagcttcag cacagctttt
     1861 ggaggagcag ctggtgtggc ggccgcagcg aacaccaggc agaagatgta gctcagggca
     1921 atgatggtct tcatggtggc gctagcaggc gcgttttcga tagaaccgag gtgcagttgg
     1981 acctgggagt ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc
     2041 taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag
     2101 ccaagggggt gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa
     2161 tccgcgttcc aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat
     2221 ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc
     2281 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     2341 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     2401 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     2461 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     2521 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     2581 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     2641 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     2701 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     2761 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     2821 taattgatta ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac
     2881 atgagccaat ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc
     2941 caatattgat ttatgctata taaccaatga ataatatggc taatggccaa tattgagtca
     3001 acgccactgt gcatgaagct cgtaattgtt atccgctcac aattaagggc gacacaaaat
     3061 ttattctaaa tgataataaa tactgataac atcttatagt ttgtattata ttttgtatta
     3121 tcgttgacat gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt
     3181 tattttctta attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa
     3241 tagatgaata gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa
     3301 gtgcgttgct tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac
     3361 aggcgccctt aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc
     3421 gccgaagcgg gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca
     3481 gggggcccga gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc
     3541 aaaaaggcca tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc
     3601 gccttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4201 ggctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4381 tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa
     4441 gggattttgg tcatgagcaa ggggtgttat gagccatatt caggtataaa tgggctcgcg
     4501 ataatgttca gaattggtta attggttgta acactgaccc ctatttgttt atttttctaa
     4561 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4621 tgaaaaagga agaatatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4681 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4741 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4801 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4861 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4921 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4981 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5041 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5101 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5161 cgtgacacca cgatgcctgt agcgatggca acaacgttgc gcaaactatt aactggcgaa
     5221 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5281 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atccggagcc
     5341 ggtgagcgtg gttctcgcgg tatcatcgca gcgctggggc cagatggtaa gccctcccgt
     5401 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5461 gctgagatag gtgcctcact gattaagcat tggtaatgac catgattacg ccaagctcta
     5521 gctagaggtc gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca
     5581 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     5641 gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc
     5701 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
     5761 atagaaggcg gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt
     5821 catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag
     5881 ccgctcgtag aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc
     5941 gcgctcggcc gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg
     6001 gtcgggcgag acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc
     6061 caggaggcct tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg
     6121 cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc
     6181 caccgcggcg ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag
     6241 gaagagttct tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg
     6301 cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc
     6361 gcgggtggcg aggcgcaccg tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc
     6421 gtattatact atgccgatat actatgccga tgattaattg tcaacacgtg ctgatcagat
     6481 ccgaaaatgg atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag
     6541 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
     6601 aaaaaaatta gtcagccatg gggcggagaa t
//
