LOCUS       sequence-393177-        8147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express single-chain pMHC with sequencing tag for 10x genomics.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8147)
  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 8147)
  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: plentiCMV10x-sct-pp65-b2m-HLA-A0201
FEATURES             Location/Qualifiers
     source          1..8147
                     /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      3566..3575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    5057..5645
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5110..5130)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5528..5539)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5717..5950
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6028..6149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6065..6084)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6119..6138
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6189..6324
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6344..6363)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6345..6363)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6373..6390)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6373..6389)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6382..6404)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6531..6986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6618..6637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6828..6849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7012..7116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7117..7185,7186..7977)
                     /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(7335..7354)
                     /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 ggagaagcag
     3541 tggtatcaac gcagagtact gtacagccac catggcgacg ggttcaagaa cttccctact
     3601 tcttgcattt ggcctgcttt gtttgccgtg gttacaggag ggctcggcaa acctcgtccc
     3661 tatggttgca actgtcggat gcggagggtc cggcggggga ggtagcggtg gtggaggaag
     3721 catccagcgt actccaaaga ttcaggttta ctcacgtcat ccagcagaga atggaaagtc
     3781 aaatttcctg aattgctatg tgtctgggtt tcatccatcc gacattgaag ttgacttact
     3841 gaagaatgga gagagaattg aaaaagtgga gcattcagac ttgtctttca gcaaggactg
     3901 gtctttctat ctcttgtact acactgaatt cacccccact gaaaaagatg agtatgcctg
     3961 ccgtgtgaac catgtgactt tgtcacagcc caagatagtt aagtgggatc gagacatggg
     4021 aggtggtggc agcggtggag ggggctccgg cggtgggtcc catataagaa acggatccgg
     4081 cggcggcggc ggaagcggct ctcactccat gaggtatttc ttcacatccg tgtcccggcc
     4141 cggccgtggg gagccccgct tcatcgcagt gggctacgtg gacgacacgc agttcgtgcg
     4201 gttcgacagc gacgccgcga gccagaggat ggagccgcgg gcgccgtgga tagagcagga
     4261 gggtccggag tattgggacg gggagacacg gaaagtgaag gcccactcac agactcaccg
     4321 agtggacctg gggaccctgc gcggctgcta caaccagagc gaggccggtt ctcacaccgt
     4381 ccagaggatg tatggctgcg acgtggggtc ggactggcgc ttcctccgcg ggtaccacca
     4441 gtacgcctac gacggcaagg attacatcgc cctgaaagag gacctgcgct cttggaccgc
     4501 ggcggacatg gcagctcaga ccaccaagca caagtgggag gcggcccatg tggcggagca
     4561 gttgagagcc tacctggagg gcacgtgcgt ggagtggctc cgcagatacc tggagaacgg
     4621 gaaagaaaca ctgcagcgca cggacgcccc caaaacgcat atgactcacc acgctgtctc
     4681 tgaccatgaa gccaccctga ggtgctgggc cctgagcttc taccctgcgg agatcacact
     4741 gacctggcag cgggatgggg aggaccagac ccaggacacg gagctcgtgg agaccaggcc
     4801 tgcaggggat ggaaccttcc agaagtgggc ggctgtggtg gtgccttctg gacaggagca
     4861 gagatacacc tgccatgtgc agcatgaggg tttgcccaag cccctcaccc tgagatggga
     4921 gccgtcttcc cagcccacca tccccatcgt gggcatcatt gctggcctgg ttctctttgg
     4981 agctgtgatc actggagctg tggtcgctgc tgtgatgtgg aggaggaaga gctcaggttg
     5041 aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5101 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5161 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5221 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5281 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5341 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5401 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     5461 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5521 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5581 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5641 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     5701 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     5761 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5821 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5881 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5941 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     6001 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6061 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6121 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6241 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6301 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     6361 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     6421 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     6481 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     6541 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6601 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6661 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6721 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6781 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6841 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     6901 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6961 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     7021 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7081 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7141 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7201 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7261 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7321 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7381 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7441 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7501 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7561 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7621 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7681 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7741 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7801 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7861 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7921 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7981 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8041 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8101 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
