LOCUS       sequence-487874-        6260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTwist CMV BetaGlobin encoding avelumab heavy chain with knob Fc and
            6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6260)
  AUTHORS   Stark JC, Gray MA, Ibarlucea-Benitez I, Lustig M, Bond A, Cho B, 
            Govil I, Luu T, Priestley MJ, Veth TS, Errington WJ, Bruncsics B, 
            Ribi MK, Williams LA, Sarkar CA, Wisnovsky S, Riley NM, Morrissey 
            MA, Valerius T, Ravetch JV, Bertozzi CR.
  TITLE     Antibody-lectin chimeras for glyco-immune checkpoint blockade.
  JOURNAL   Nat Biotechnol 2025
REFERENCE   2  (bases 1 to 6260)
  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"; journalName: "Nat 
            Biotechnol 2025"
COMMENT     Sequence Label: pTCB-avelHCknob6xHis
FEATURES             Location/Qualifiers
     source          1..6260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1414..1431
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1484..1605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1495..1514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1549..1568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1720..1766
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1796..1932
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1977..2048
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2274..2862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2354..2373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3608..3676,3677..4468)
                     /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(3826..3845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4481..4617
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4769..4840
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4845..4891)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4909..4957
                     /note="synthetic polyadenylation signal"
     enhancer        5038..5341
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5343..5545
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5496..5516
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          5679..6154
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
ORIGIN
        1 ctcgagatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
       61 tccgaagttc aattgctgga gagcggcgga ggtttggtcc agcccggcgg ttctttgcga
      121 ctgtcctgcg cagcctctgg tttcacattt tcttcctaca ttatgatgtg ggtgcgccag
      181 gctcctggaa agggacttga atgggtaagc agcatttatc cctctggtgg gatcactttc
      241 tacgccgata ccgtgaaggg gagattcact atatcacgag acaactcaaa gaataccctc
      301 tatcttcaga tgaatagttt gcgagccgag gatacggctg tctactactg tgccaggatc
      361 aagttgggta ccgtgacaac tgttgattat tggggtcaag gaactctcgt aaccgtcagc
      421 agtgctagca ccaagggccc atcggtcttc cccctggcac cctcctccaa gagcacctct
      481 gggggcacag cggccctggg ctgcctggtc aaggactact tccccgaacc ggtgacggtg
      541 tcgtggaact caggcgccct gaccagcggc gtgcacacct tcccggctgt cctacagtcc
      601 tcaggactct actccctcag cagcgtggtg accgtgccct ccagcagctt gggcacccag
      661 acctacatct gcaacgtgaa tcacaagccc agcaacacca aggtggacaa gaaagttgag
      721 cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga actcctgggg
      781 ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc
      841 cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac
      901 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac
      961 aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc
     1021 aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga gaaaaccatc
     1081 tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atgccgggat
     1141 gagctgacca agaaccaggt cagcctgtgg tgcctggtca aaggcttcta tcccagcgac
     1201 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc
     1261 gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg
     1321 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac
     1381 acgcagaaga gcctctccct gtctccgggt gggcatcacc accatcacca ttgagctagc
     1441 ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag
     1501 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1561 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1621 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     1681 ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct
     1741 ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt
     1801 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     1861 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     1921 atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt
     1981 cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc
     2041 cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc
     2101 tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt
     2161 cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga
     2221 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2281 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2341 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2401 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2461 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2521 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2581 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2641 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2701 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2761 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2821 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2881 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2941 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3001 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3061 atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca
     3121 aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg
     3181 aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct
     3241 atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca
     3301 acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc
     3361 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     3421 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     3481 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     3541 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     3601 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3661 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3721 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3781 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3841 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3901 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3961 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4021 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4081 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4141 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     4201 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     4261 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4321 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4381 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4441 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     4501 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     4561 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     4621 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     4681 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     4741 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     4801 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     4861 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     4921 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     4981 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     5041 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     5101 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     5161 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     5221 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     5281 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     5341 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     5401 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     5461 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     5521 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcgcct ggagacgcca
     5581 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc cctcgaagct
     5641 tacatgtggt accgagctcg gatcctgaga acttcagggt gagtctatgg gacccttgat
     5701 gttttctttc cccttctttt ctatggttaa gttcatgtca taggaagggg agaagtaaca
     5761 gggtacacat attgaccaaa tcagggtaat tttgcatttg taattttaaa aaatgctttc
     5821 ttcttttaat atactttttt gtttatctta tttctaatac tttccctaat ctctttcttt
     5881 cagggcaata atgatacaat gtatcatgcc tctttgcacc attctaaaga ataacagtga
     5941 taatttctgg gttaaggcaa tagcaatatt tctgcatata aatatttctg catataaatt
     6001 gtaactgatg taagaggttt catattgcta atagcagcta caatccagct accattctgc
     6061 ttttatttta tggttgggat aaggctggat tattctgagt ccaagctagg cccttttgct
     6121 aatcatgttc atacctctta tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt
     6181 gctggcccat cactttggca aaggcggccg ctgccaagct tccgagctct cgaattcaaa
     6241 ggaggtaccg cgatatctac
//
