LOCUS       sequence-488037-        6269 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  pTwist CMV BetaGlobin encoding pembrolizumab heavy chain with LALA 
            P329G knob Fc and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6269)
  AUTHORS   Stark JC, Gray MA, Wisnovsky S, Ibarlucea-Benitez I, Lustig M, Riley
            NM, Ribi MK, Errington WJ, Bruncsics B, Sarkar CA, Valerius TV, 
            Ravetch JV, Bertozzi CR
  TITLE     Antibody-lectin chimeras for glyco-immune checkpoint blockade
  JOURNAL   bioRxiv 2022.10.26.513931; doi: 
            https://doi.org/10.1101/2022.10.26.513931
REFERENCE   2  (bases 1 to 6269)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2022.10.26.513931; doi: https://doi.org/10.1101/2022.10.26.513931"
COMMENT     Sequence Label: pTCB-pembroHCLALAPGknobHA
FEATURES             Location/Qualifiers
     source          1..6269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1414..1440
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1493..1614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1504..1523)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1558..1577
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1729..1775
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1805..1941
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1986..2057
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2283..2871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2363..2382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3617..3685,3686..4477)
                     /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(3835..3854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4490..4626
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4778..4849
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4854..4900)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4918..4966
                     /note="synthetic polyadenylation signal"
     enhancer        5047..5350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5352..5554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5505..5525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          5688..6163
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
ORIGIN
        1 ctcgagatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
       61 tcccaagtgc aattggtgca gtccggcgtt gaagttaaaa agcccggtgc gtccgtgaag
      121 gtttcctgta aggcgagtgg atacaccttc acaaactatt atatgtattg ggtcaggcaa
      181 gctccgggac agggtttgga atggatggga ggtatcaatc catccaatgg tggtacgaat
      241 tttaatgaga agttcaaaaa tagagtgact ctgactaccg acagttctac gactacagcc
      301 tacatggaat tgaagtccct gcaattcgac gatacggcag tttattattg tgctaggagg
      361 gattatagat tcgacatggg ttttgattat tggggccaag ggactactgt aacagtatca
      421 tccgctagca 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 agcagccggg
      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 gccctcggag 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 gggtacccat acgatgttcc agattacgct
     1441 tgagctagct tgactgactg agatacagcg taccttcagc tcacagacat gataagatac
     1501 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1561 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1621 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     1681 aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc
     1741 ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg attgctatct
     1801 accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     1861 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     1921 ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct cgatttcgat
     1981 caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa
     2041 actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta attaaccagt
     2101 caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca gaattgcgtc
     2161 agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag tttcatttaa
     2221 atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2281 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2341 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2401 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2461 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2521 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2581 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2641 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2701 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2761 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2821 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2881 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2941 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3001 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3061 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt taaatttccg
     3121 aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca tcttgcaggc
     3181 tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg ctattgcttg
     3241 gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg atcacccgag
     3301 agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat atcgaaatcg
     3361 gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt
     3421 tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata
     3481 ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc
     3541 tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc
     3601 agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3661 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3721 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3781 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     3841 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3901 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     3961 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4021 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4081 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4141 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     4201 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     4261 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4321 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4381 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4441 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     4501 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     4561 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     4621 ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     4681 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     4741 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     4801 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     4861 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     4921 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     4981 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5041 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     5101 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     5161 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     5221 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     5281 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     5341 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     5401 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     5461 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     5521 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcgcctg
     5581 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccc
     5641 ctcgaagctt acatgtggta ccgagctcgg atcctgagaa cttcagggtg agtctatggg
     5701 acccttgatg ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga
     5761 gaagtaacag ggtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa
     5821 aatgctttct tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc
     5881 tctttctttc agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa
     5941 taacagtgat aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc
     6001 atataaattg taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta
     6061 ccattctgct tttattttat ggttgggata aggctggatt attctgagtc caagctaggc
     6121 ccttttgcta atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct
     6181 ggtctgtgtg ctggcccatc actttggcaa aggcggccgc tgccaagctt ccgagctctc
     6241 gaattcaaag gaggtaccgc gatatctac
//
