LOCUS       sequence-488045-        5546 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  pTwist CMV BetaGlobin encoding pembrolizumab light chain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5546)
  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 5546)
  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-pembroLC
FEATURES             Location/Qualifiers
     source          1..5546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    770..891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(781..800)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     835..854
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1006..1052
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1082..1218
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1263..1334
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1560..2148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1640..1659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2894..2962,2963..3754)
                     /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(3112..3131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3767..3903
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4055..4126
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4131..4177)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4195..4243
                     /note="synthetic polyadenylation signal"
     enhancer        4324..4627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4629..4831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4782..4802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          4965..5440
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
ORIGIN
        1 ctcgagatgg gatggtcatg tatcatcctt tttctagtag caactgcaac cggtgtacat
       61 tccgaaattg ttctcactca gtctccggca acgttgtcac tcagtcctgg tgagagagct
      121 acgttgtcct gccgggctag caagggagtt tcaacatcag gatattccta cctccactgg
      181 taccaacaga aaccgggtca ggctccgcgc ctcctcattt acctcgcatc ctacctggaa
      241 tctggggtgc ccgcgcggtt ttctgggtct ggctctggga ctgatttcac tctcactatt
      301 tcctcattgg aacccgagga ttttgcggtg tactattgcc agcactcacg ggatctgcct
      361 cttacttttg gaggaggtac taaagtggaa ataaaacgca ccgtcgccgc gcccagcgtg
      421 ttcatcttcc cgcccagcga cgagcagctg aagagcggca cggccagcgt ggtctgcctg
      481 ctcaacaact tctacccccg ggaggccaag gtgcagtgga aggtggacaa cgccctgcag
      541 tcggggaaca gccaggagag cgtcaccgag caggacagca aggacagcac ctacagcctg
      601 tcgagcaccc tcacgctgag caaggccgac tacgagaagc acaaggtgta cgcgtgcgag
      661 gtgacccacc agggcctgag cagccccgtc accaagtcgt tcaaccgcgg agaatgctga
      721 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
      781 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
      841 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
      901 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
      961 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     1021 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     1081 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     1141 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     1201 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     1261 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     1321 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     1381 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     1441 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     1501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1981 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     2401 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     2461 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     2521 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     2581 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     2641 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     2701 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     2761 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     2821 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     2881 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2941 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3001 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3061 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     3121 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3181 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     3241 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3301 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3361 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3421 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     3481 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     3541 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3601 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3661 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3721 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     3781 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     3841 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     3901 taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     3961 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     4021 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     4081 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     4141 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
     4201 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     4261 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     4321 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     4381 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     4441 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     4501 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     4561 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     4621 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     4681 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     4741 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     4801 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcgcctggag
     4861 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctcccctc
     4921 gaagcttaca tgtggtaccg agctcggatc ctgagaactt cagggtgagt ctatgggacc
     4981 cttgatgttt tctttcccct tcttttctat ggttaagttc atgtcatagg aaggggagaa
     5041 gtaacagggt acacatattg accaaatcag ggtaattttg catttgtaat tttaaaaaat
     5101 gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct
     5161 ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa
     5221 cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata
     5281 taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca
     5341 ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct
     5401 tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt
     5461 ctgtgtgctg gcccatcact ttggcaaagg cggccgctgc caagcttccg agctctcgaa
     5521 ttcaaaggag gtaccgcgat atctac
//
