LOCUS       sequence-386251-        8104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express single-chain peptide-MHC (pp65 CMV epitope on HLA-A0201).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8104)
  AUTHORS   Yu B, Shi Q, Belk JA, Yost KE, Parker KR, Li R, Liu BB, Huang H, 
            Lingwood D, Greenleaf WJ, Davis MM, Satpathy AT, Chang HY
  TITLE     Engineered cell entry links receptor biology with single-cell 
            genomics.
  JOURNAL   Cell. 2022 Dec 22;185(26):4904-4920.e22. doi: 
            10.1016/j.cell.2022.11.016. Epub 2022 Dec 13.
  PUBMED    36516854
REFERENCE   2  (bases 1 to 8104)
  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"; doi: 
            "10.1016/j.cell.2022.11.016"; journalName: "Cell"; date: 
            "2022-12-22- 22"; volume: "185"; issue: "26"; pages: "4904-4920.e22"
COMMENT     Sequence Label: plentiCMV-sct-pp65-b2m-HLA-A0201
FEATURES             Location/Qualifiers
     source          1..8104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2859..2975
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2998..3301
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3302..3505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3456..3476
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3502..3526
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3541..3550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    5014..5602
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5067..5087)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5485..5496)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5674..5907
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5985..6106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6022..6041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6076..6095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6146..6281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6301..6320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6302..6320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6330..6347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6330..6346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6339..6361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6488..6943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6575..6594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6785..6806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6969..7073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7074..7142,7143..7934)
                     /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(7292..7311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2821 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     2881 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2941 gaattacaaa aacaaattac aaaattcaaa attttatcga taagcttggg agttccgcgt
     3001 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3061 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     3121 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     3181 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     3241 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3301 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3361 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     3421 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     3481 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagtgtaca
     3541 gccaccatgg cgacgggttc aagaacttcc ctacttcttg catttggcct gctttgtttg
     3601 ccgtggttac aggagggctc ggcaaacctc gtccctatgg ttgcaactgt cggatgcgga
     3661 gggtccggcg ggggaggtag cggtggtgga ggaagcatcc agcgtactcc aaagattcag
     3721 gtttactcac gtcatccagc agagaatgga aagtcaaatt tcctgaattg ctatgtgtct
     3781 gggtttcatc catccgacat tgaagttgac ttactgaaga atggagagag aattgaaaaa
     3841 gtggagcatt cagacttgtc tttcagcaag gactggtctt tctatctctt gtactacact
     3901 gaattcaccc ccactgaaaa agatgagtat gcctgccgtg tgaaccatgt gactttgtca
     3961 cagcccaaga tagttaagtg ggatcgagac atgggaggtg gtggcagcgg tggagggggc
     4021 tccggcggtg ggggatccgg cggcggcgga agcggctctc actccatgag gtatttcttc
     4081 acatccgtgt cccggcccgg ccgtggggag ccccgcttca tcgcagtggg ctacgtggac
     4141 gacacgcagt tcgtgcggtt cgacagcgac gccgcgagcc agaggatgga gccgcgggcg
     4201 ccgtggatag agcaggaggg tccggagtat tgggacgggg agacacggaa agtgaaggcc
     4261 cactcacaga ctcaccgagt ggacctgggg accctgcgcg gctgctacaa ccagagcgag
     4321 gccggttctc acaccgtcca gaggatgtat ggctgcgacg tggggtcgga ctggcgcttc
     4381 ctccgcgggt accaccagta cgcctacgac ggcaaggatt acatcgccct gaaagaggac
     4441 ctgcgctctt ggaccgcggc ggacatggca gctcagacca ccaagcacaa gtgggaggcg
     4501 gcccatgtgg cggagcagtt gagagcctac ctggagggca cgtgcgtgga gtggctccgc
     4561 agatacctgg agaacgggaa agaaacactg cagcgcacgg acgcccccaa aacgcatatg
     4621 actcaccacg ctgtctctga ccatgaagcc accctgaggt gctgggccct gagcttctac
     4681 cctgcggaga tcacactgac ctggcagcgg gatggggagg accagaccca ggacacggag
     4741 ctcgtggaga ccaggcctgc aggggatgga accttccaga agtgggcggc tgtggtggtg
     4801 ccttctggac aggagcagag atacacctgc catgtgcagc atgagggttt gcccaagccc
     4861 ctcaccctga gatgggagcc gtcttcccag cccaccatcc ccatcgtggg catcattgct
     4921 ggcctggttc tctttggagc tgtgatcact ggagctgtgg tcgctgctgt gatgtggagg
     4981 aggaagagct caggttgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     5041 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5101 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5161 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5221 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5281 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5341 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     5401 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     5461 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     5521 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     5581 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     5641 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5701 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5761 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5821 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5881 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
     5941 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6001 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6061 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6121 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6181 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6241 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     6301 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     6361 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6421 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6481 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6541 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6601 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6661 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6721 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6781 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6841 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6901 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     6961 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8101 cttc
//
