LOCUS       sequence-368535-        6601 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 6601)
  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 6601)
  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-TNFRSF13B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..128
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(147..194)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(242..571)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      256..391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(310..329)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     556..576
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    694..805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     794..811
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(951..968)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(969..1013)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2325..2344)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2357..2375)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2358..2375)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2585..2609)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2606..2809)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2635..2655)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2810..3189
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3356..3372
                     /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(3828..3845)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3828..3844)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3943..3960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4114..4702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4194..4213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4788..4807)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4813..4832)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4874..4965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4966..5034,5035..5826)
                     /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(5184..5203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5879..6000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5890..5909)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5944..5963
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6130..6601,1..128))
                     /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(6213..6233)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 atgtggcggt ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg
       61 agggtgcgta cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac
      121 tcggtcatgg tttagttcct caccttgtcg tattatacta tgccgatata ctatgccgat
      181 gattaattgt caacacgtgc tgatcagatc cgaaaatgga tatacaagct cccgggagct
      241 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
      301 cagaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
      361 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
      421 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      481 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      541 ctggggactt tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt
      601 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      661 aaaagaatag accgagatag ggttgagtgt tgtgatgcaa tttcctcatt ttattaggaa
      721 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
      781 gatggctggc aactagaagg cacagtcgag gctgattttc tctatctatt tattttacta
      841 ctcaagacta tcttgcaaag tagaagaacc ctgtctgcag tatgacgacc gtgtataccc
      901 gattggctta ccgggtaacg cgttgtgcca gtgactacag gccggcctta gtggtgatgg
      961 tggtggtgtt catgccactc aatcttctga gcctcgaaaa tatcattaag accgctgccg
     1021 ccggggctca ggctcaggga cttctgggtg tagtggttgt gcagggcctc gtgcatcacg
     1081 ctgcagctga acacgttgcc ctgctgccac ctggacttgt ccacggtcag cttgctgtac
     1141 aggaagaagc tgccgtcgct gtccagcact gggggggtgg tcttgtagtt gttctcgggc
     1201 tggccgttgc tctcccactc cacggcgata tcgctggggt agaagccctt caccagacag
     1261 gtcagggaca cctggttctt ggtcatctcc tcccggctgg ggggcagggt gtacacctgg
     1321 ggctcccgtg gctggccctt ggccttgctg attgtctttt cgattggggc agccagggcc
     1381 ttgttggaga ctttgcactt gtattctttg ccgttcagcc agtcctggtg cagcacggtc
     1441 agcacggaca ccaccctgta ggtgctgttg tactgctcct ctctgggctt ggtcttggcg
     1501 ttgtgcacct ccacgccgtc cacgtaccag ttgaacttca cctctgggtc ctcgtgggac
     1561 acggccacca ccacgcaggt cacctcgggg gtcctgctga tcatcagggt gtccttgggc
     1621 ttggggggga acaggaacac ggaaggccct cccagcagtt ctggagctgg gcaggggggg
     1681 caggtgtggg tcttgtcgcc gcccggcgcg ccagagtaca ccagggccac ctgatcagca
     1741 gacagcttca ggccaggcag agcaggagaa gcttcagatc ctctgtgctc gaggccctgg
     1801 tatctgccgg agttgtcgct gttgttttcc acctcgccgc ttctctgcct tctcagttca
     1861 ggaggcaggt tcacggggct tctcagcttg ttctcgcaga agtaggcgca ctgcttaggg
     1921 tgctgtccgc aaatagaggc gcagctgatg caatcccgca gcagatgatc gtagaacttg
     1981 ccctgctctt ttctacagct caggcttcta cagaaggcgg cgcaggttct ctggctctgg
     2041 tggttgcaga tggtcttgca gctcatacag gtgcccagca gagggtccca gtactgttcc
     2101 tcaggacaag atctcatggc cacgccggtc cacaggcctt gagggaatct ttcctcttgg
     2161 tccactctgg atctgccgcc tcttcttgat cttcccagtc cagacatggc ggccgcagcg
     2221 aacaccaggc agaagatgta gctcagggca atgatggtct tcatggtggc gctagcaggc
     2281 gcgttttcga tagaaccgag gtgcagttgg acctgggagt ggacacctgt ggagagaaag
     2341 gcaaagtgga tgttattcta tagtgtcacc taaatcgtat gtgtatgata cataaggtta
     2401 tgtattaatt gtagccgcgt tctaacgaag ccaagggggt gggcctatag actctatagg
     2461 cggtacttac gtcactcttg gcacggggaa tccgcgttcc aatgcaccgt tcccggccgc
     2521 ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc
     2581 aggcgatctg acggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc
     2641 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     2701 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     2761 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
     2821 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     2881 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     2941 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     3001 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     3061 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     3121 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     3181 atcaatgcca acatggcggt catattggac atgagccaat ataaatgtac atattatgat
     3241 atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata taaccaatga
     3301 ataatatggc taatggccaa tattgagtca acgccactgt gcatgaagct cgtaattgtt
     3361 atccgctcac aattaagggc gacacaaaat ttattctaaa tgataataaa tactgataac
     3421 atcttatagt ttgtattata ttttgtatta tcgttgacat gtataatttt gatatcaaaa
     3481 actgattttc cctttattat tttcgagatt tattttctta attctcttta acaaactaga
     3541 aatattgtat atacaaaaaa tcataaataa tagatgaata gtttaattat aggtgttcat
     3601 caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct tttttctcat ttataaggtt
     3661 aaataattct catatatcaa gcaaagtgac aggcgccctt aaatattctg acaaatgctc
     3721 tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga
     3781 ttaacgatta ctcgttatca gaaccgccca gggggcccga gcttaagact ggccgtcgtt
     3841 ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca tccgtcaggg gccttctgct
     3901 tagtttgatg cctggcagtt ccctactctc gccttccgct tcctcgctca ctgactcgct
     3961 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4021 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4081 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4141 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4201 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4261 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4321 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4381 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4441 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4501 aggcggtgct acagagttct tgaagtggtg ggctaactac ggctacacta gaagaacagt
     4561 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4621 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4681 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4741 gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg tcatgagcaa ggggtgttat
     4801 gagccatatt caggtataaa tgggctcgcg ataatgttca gaattggtta attggttgta
     4861 acactgaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4921 caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag tattcaacat
     4981 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5041 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5101 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5161 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5221 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5281 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5341 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5401 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5461 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcgatggca
     5521 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5581 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5641 ggctggttta ttgctgataa atccggagcc ggtgagcgtg gttctcgcgg tatcatcgca
     5701 gcgctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5761 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5821 tggtaatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata
     5881 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     5941 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     6001 ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     6061 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     6121 tagcacgtgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac
     6181 ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc
     6241 ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac
     6301 gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg tggccaggaa
     6361 ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc
     6421 cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc
     6481 ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac
     6541 cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc
     6601 g
//
