LOCUS       sequence-487875-        6251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTwist CMV BetaGlobin encoding magrolimab heavy chain with LALA 
            P329G knob Fc and 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6251)
  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 6251)
  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-magroHCLALAPGknob6xHis
FEATURES             Location/Qualifiers
     source          1..6251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1405..1422
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1475..1596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1486..1505)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1540..1559
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1711..1757
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1787..1923
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1968..2039
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2265..2853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2345..2364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3599..3667,3668..4459)
                     /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(3817..3836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4472..4608
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4760..4831
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4836..4882)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4900..4948
                     /note="synthetic polyadenylation signal"
     enhancer        5029..5332
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5334..5536
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5487..5507
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          5670..6145
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
ORIGIN
        1 ctcgagatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
       61 tcccaagttc aacttcaaca acccggcgca gagctggtta agcctggggc gagtgtaatg
      121 atgagttgca aagcaagcgg atacacgttt accaactaca atatgcattg ggtaaaacag
      181 acgcctgggc aaggtcttga atggatcggc accatctacc ctggcaacga tgatacgtca
      241 tacaaccaga agtttaagga taaggccacg ctgaccgccg ataaatcctc ctctgcagcg
      301 tacatgcagt tgtctagtct tacgtctgag gactctgcgg tgtactattg cgccaggggg
      361 ggttatagag ctatggacta ctgggggcaa gggacctccg tgaccgtatc ctcagctagc
      421 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca
      481 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac
      541 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc
      601 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc
      661 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct
      721 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagcagccgg gggaccgtca
      781 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc
      841 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg
      901 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg
      961 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac
     1021 aagtgcaagg tctccaacaa agccctcgga gcccccatcg agaaaaccat ctccaaagcc
     1081 aaagggcagc cccgagaacc acaggtgtac accctgcccc catgccggga tgagctgacc
     1141 aagaaccagg tcagcctgtg gtgcctggtc aaaggcttct atcccagcga catcgccgtg
     1201 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac
     1261 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag
     1321 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag
     1381 agcctctccc tgtctccggg tgggcatcac caccatcacc attgagctag cttgactgac
     1441 tgagatacag cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa
     1501 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1561 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1621 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1681 tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg
     1741 tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa
     1801 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     1861 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     1921 taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg
     1981 gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa
     2041 tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg
     2101 agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt
     2161 ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc
     2221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3061 atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg
     3121 tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta
     3181 tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg
     3241 tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc
     3301 ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc
     3361 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     3421 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     3481 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     3541 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     3601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     3721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3781 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3901 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     4021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4141 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     4201 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4441 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     4501 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     4561 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     4621 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     4681 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     4741 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     4801 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     4861 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     4921 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     4981 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5041 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5101 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     5161 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     5221 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     5281 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
     5341 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     5401 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     5461 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     5521 ctatataagc agagctggtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     5581 ttttgacctc catagaagac accgggaccg atccagcctc ccctcgaagc ttacatgtgg
     5641 taccgagctc ggatcctgag aacttcaggg tgagtctatg ggacccttga tgttttcttt
     5701 ccccttcttt tctatggtta agttcatgtc ataggaaggg gagaagtaac agggtacaca
     5761 tattgaccaa atcagggtaa ttttgcattt gtaattttaa aaaatgcttt cttcttttaa
     5821 tatacttttt tgtttatctt atttctaata ctttccctaa tctctttctt tcagggcaat
     5881 aatgatacaa tgtatcatgc ctctttgcac cattctaaag aataacagtg ataatttctg
     5941 ggttaaggca atagcaatat ttctgcatat aaatatttct gcatataaat tgtaactgat
     6001 gtaagaggtt tcatattgct aatagcagct acaatccagc taccattctg cttttatttt
     6061 atggttggga taaggctgga ttattctgag tccaagctag gcccttttgc taatcatgtt
     6121 catacctctt atcttcctcc cacagctcct gggcaacgtg ctggtctgtg tgctggccca
     6181 tcactttggc aaaggcggcc gctgccaagc ttccgagctc tcgaattcaa aggaggtacc
     6241 gcgatatcta c
//
