LOCUS       sequence-487856-        6014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTwist CMV BetaGlobin encoding Galectin-9 N terminal carbohydrate 
            binding domain-Fc fusion with LALA P329G hole Fc and 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  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 6014)
  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-Gal9NFcLALAPGhole6xHis
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1168..1185
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1238..1359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1249..1268)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1303..1322
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1474..1520
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1550..1686
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1731..1802
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2028..2616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2108..2127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3362..3430,3431..4222)
                     /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(3580..3599)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4235..4371
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4523..4594
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4599..4645)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4663..4711
                     /note="synthetic polyadenylation signal"
     enhancer        4792..5095
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5097..5299
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5250..5270
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          5433..5908
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
ORIGIN
        1 ctcgagatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
       61 tccttttctg ggactattca aggaggtctc caggacggac ttcagatcac tgtcaatggg
      121 accgttctca gctccagtgg aaccaggttt gctgtgaact ttcagactgg cttcagtgga
      181 aatgacattg ccttccactt caaccctcgg tttgaagatg gagggtacgt ggtgtgcaac
      241 acgaggcaga acggaagctg ggggcccgag gagaggaaga cacacatgcc tttccagaag
      301 gggatgccct ttgacctctg cttcctggtg cagagctcag atttcaaggt gatggtgaac
      361 gggatcctct tcgtgcagta cttccaccgc gtgcccttcc accgtgtgga caccatctcc
      421 gtcaatggct ctgtgcagct gtcctacatc agcttccagg gcggaggtgg aggccccaaa
      481 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaagcagc cgggggaccg
      541 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag
      601 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac
      661 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc
      721 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag
      781 tacaagtgca aggtctccaa caaagccctc ggagccccca tcgagaaaac catctccaaa
      841 gccaaagggc agccccgaga accacaggtg tgcaccctgc ccccaagccg ggatgagctg
      901 accaagaacc aggtcagcct gagctgcgcg gtcaaaggct tctatcccag cgacatcgcc
      961 gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg
     1021 gactccgacg gctccttctt cctcgtgagc aagctcaccg tggacaagag caggtggcag
     1081 caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag
     1141 aagagcctct ccctgtctcc gggtgggcat caccaccatc accattgagc tagcttgact
     1201 gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga
     1261 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1321 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1381 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1441 aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac
     1501 gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca
     1561 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     1621 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     1681 ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta
     1741 atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc
     1801 taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg
     1861 gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct
     1921 ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa
     1981 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2041 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2101 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2161 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2221 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2281 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2341 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2401 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2461 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2521 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2581 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2641 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2701 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2761 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2821 gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc
     2881 tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa
     2941 ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc
     3001 aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac
     3061 atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt
     3121 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     3181 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     3241 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     3301 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     3361 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3421 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3481 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3541 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3601 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3661 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3721 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3781 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3841 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3901 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     3961 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4021 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4081 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4141 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4201 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     4261 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     4321 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa
     4381 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     4441 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     4501 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     4561 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     4621 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     4681 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     4741 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     4801 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     4861 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     4921 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     4981 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     5041 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     5101 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     5161 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     5221 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     5281 ggtctatata agcagagctg gtttagtgaa ccgtcagatc gcctggagac gccatccacg
     5341 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctcccctcga agcttacatg
     5401 tggtaccgag ctcggatcct gagaacttca gggtgagtct atgggaccct tgatgttttc
     5461 tttccccttc ttttctatgg ttaagttcat gtcataggaa ggggagaagt aacagggtac
     5521 acatattgac caaatcaggg taattttgca tttgtaattt taaaaaatgc tttcttcttt
     5581 taatatactt ttttgtttat cttatttcta atactttccc taatctcttt ctttcagggc
     5641 aataatgata caatgtatca tgcctctttg caccattcta aagaataaca gtgataattt
     5701 ctgggttaag gcaatagcaa tatttctgca tataaatatt tctgcatata aattgtaact
     5761 gatgtaagag gtttcatatt gctaatagca gctacaatcc agctaccatt ctgcttttat
     5821 tttatggttg ggataaggct ggattattct gagtccaagc taggcccttt tgctaatcat
     5881 gttcatacct cttatcttcc tcccacagct cctgggcaac gtgctggtct gtgtgctggc
     5941 ccatcacttt ggcaaaggcg gccgctgcca agcttccgag ctctcgaatt caaaggaggt
     6001 accgcgatat ctac
//
