LOCUS       sequence-490226-        4841 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Computationally designed water-soluble mimic of the transmembrane 
            protein CD20 dimer. For expression as secreted protein in Expi293 
            cell. Insert: HSA-his8-TEVsolubleCD20- (10.4kDa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4841)
  AUTHORS   Yao Z, Kuhlman B
  TITLE     Design of a water-soluble CD20 antigen with computational epitope 
            scaffolding.
  JOURNAL   Protein Sci. 2025 Aug;34(8):e70184. doi: 10.1002/pro.70184.
  PUBMED    40671354
REFERENCE   2  (bases 1 to 4841)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2025-08"; volume: "34(8):e70184. doi"; pages: " 
            10.1002/pro.70184"
COMMENT     Sequence Label: Soluble CD20His
FEATURES             Location/Qualifiers
     source          1..4841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1822..1845
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1870..1890
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2196..2215)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2261..2316
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2315..2334)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2677..2693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2677..2693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2690..2712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2701..2717
                     /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)."
     promoter        complement(join(2725..2731,2732..2749,2750..2755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2770..2791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2920..2937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3091..3679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3171..3190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3850..4641,4642..4710))
                     /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     4473..4492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4711..4815)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg gggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      541 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagcgc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attgcggccg
     1741 tctcaggccg agttcggtac cgccaccatg aagtgggtaa cctttatttc cttgctgttc
     1801 ctctttagct cggcttattc ccatcaccac catcatcacc atcatggcgg tgggagtggt
     1861 ggtgggagcg agaacctgta cttccaatcc cacacgcctt acataaatat ctacaattgt
     1921 gagccggcca acccctcaga gaagaactcc ccgtcaaccc agtactgtta cttggctcag
     1981 gagctgctct caaagaatag acacttggag aatgaagtga agagactcaa aaaacttgtg
     2041 gacgatttgg aagatgaact caaggcccag aaagagaagt acaaagccat tagctagtag
     2101 tgactcgagc gataattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     2161 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     2221 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     2281 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2341 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     2401 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     2461 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     2521 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     2581 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     2641 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2821 gtcgggaaac ctgtcgtgcc agcggatcga tccgctgcat taatgaatcg gccaacgcgc
     2881 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2941 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3001 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3061 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3121 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3181 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3241 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3301 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3361 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3421 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4501 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4681 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4741 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
